{"id":2499,"date":"2023-04-16T21:40:46","date_gmt":"2023-04-17T02:40:46","guid":{"rendered":"https:\/\/nuevo22.cidsamexico.com\/?page_id=2499"},"modified":"2023-04-18T14:17:53","modified_gmt":"2023-04-18T19:17:53","slug":"etiquetas-his-tag","status":"publish","type":"page","link":"https:\/\/nuevo22.cidsamexico.com\/index.php\/etiquetas-his-tag\/","title":{"rendered":"His-Tag: Fundamentos y principios"},"content":{"rendered":"\t\t<div data-elementor-type=\"wp-post\" data-elementor-id=\"2499\" class=\"elementor elementor-2499\">\n\t\t\t\t\t\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-2c888a9 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"2c888a9\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-494b2ee\" data-id=\"494b2ee\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-2fdd245 elementor-widget elementor-widget-image\" data-id=\"2fdd245\" data-element_type=\"widget\" data-widget_type=\"image.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t<style>\/*! elementor - v3.7.8 - 02-10-2022 *\/\n.elementor-widget-image{text-align:center}.elementor-widget-image a{display:inline-block}.elementor-widget-image a img[src$=\".svg\"]{width:48px}.elementor-widget-image img{vertical-align:middle;display:inline-block}<\/style>\t\t\t\t\t\t\t\t\t\t\t\t<img width=\"600\" height=\"346\" src=\"https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/His-tag-Abbildung-1-600x346.jpg\" class=\"attachment-classic-grid-listing size-classic-grid-listing\" alt=\"\" loading=\"lazy\" srcset=\"https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/His-tag-Abbildung-1-600x346.jpg 600w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/His-tag-Abbildung-1-300x173.jpg 300w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/His-tag-Abbildung-1-1024x590.jpg 1024w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/His-tag-Abbildung-1-768x443.jpg 768w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/His-tag-Abbildung-1-1536x886.jpg 1536w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/His-tag-Abbildung-1-278x160.jpg 278w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/His-tag-Abbildung-1.jpg 2026w\" sizes=\"(max-width: 600px) 100vw, 600px\" \/>\t\t\t\t\t\t\t\t\t\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-98f2b66 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"98f2b66\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-d90f0d5\" data-id=\"d90f0d5\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-cbbe079 elementor-widget elementor-widget-spacer\" data-id=\"cbbe079\" data-element_type=\"widget\" data-widget_type=\"spacer.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t<style>\/*! elementor - v3.7.8 - 02-10-2022 *\/\n.elementor-column .elementor-spacer-inner{height:var(--spacer-size)}.e-container{--container-widget-width:100%}.e-container>.elementor-widget-spacer{width:var(--container-widget-width,var(--spacer-size));-ms-flex-item-align:stretch;align-self:stretch;-ms-flex-negative:0;flex-shrink:0}.e-container>.elementor-widget-spacer>.elementor-widget-container,.e-container>.elementor-widget-spacer>.elementor-widget-container>.elementor-spacer{height:100%}.e-container>.elementor-widget-spacer>.elementor-widget-container>.elementor-spacer>.elementor-spacer-inner{height:var(--container-widget-height,var(--spacer-size))}<\/style>\t\t<div class=\"elementor-spacer\">\n\t\t\t<div class=\"elementor-spacer-inner\"><\/div>\n\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-2de07b6 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"2de07b6\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-b938d2a\" data-id=\"b938d2a\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-3a45592 elementor-widget elementor-widget-text-editor\" data-id=\"3a45592\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t<style>\/*! elementor - v3.7.8 - 02-10-2022 *\/\n.elementor-widget-text-editor.elementor-drop-cap-view-stacked .elementor-drop-cap{background-color:#818a91;color:#fff}.elementor-widget-text-editor.elementor-drop-cap-view-framed .elementor-drop-cap{color:#818a91;border:3px solid;background-color:transparent}.elementor-widget-text-editor:not(.elementor-drop-cap-view-default) .elementor-drop-cap{margin-top:8px}.elementor-widget-text-editor:not(.elementor-drop-cap-view-default) .elementor-drop-cap-letter{width:1em;height:1em}.elementor-widget-text-editor .elementor-drop-cap{float:left;text-align:center;line-height:1;font-size:50px}.elementor-widget-text-editor .elementor-drop-cap-letter{display:inline-block}<\/style>\t\t\t\t<p>La etiqueta de polihistidina o His6 es una etiqueta de prote\u00edna desarrollada originalmente para la purificaci\u00f3n eficiente de prote\u00ednas en 1988 (Hochuli, Bannwarth &amp; D\u00f6beli et al., 1988). Como resultado de su relaci\u00f3n costo-beneficio comparablemente grande, es conocida como una de las etiquetas de afinidad de prote\u00ednas m\u00e1s utilizadas hasta la fecha. Debido a su peque\u00f1o tama\u00f1o, variable entre cuatro y diez amino\u00e1cidos, rara vez interfiere con la funci\u00f3n de la prote\u00edna objtetivo. Adem\u00e1s, su baja inmunogenicidad y versatilidad en condiciones nativas y desnaturalizantes hacen que esta etiqueta de prote\u00edna se destaque entre otras. El principio de purificaci\u00f3n subyacente de la etiqueta His es una interacci\u00f3n entre los iones met\u00e1licos y el anillo de imidazol de la histidina.<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-2390cbc elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"2390cbc\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-004b86a\" data-id=\"004b86a\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-ff24a28 elementor-widget-divider--view-line elementor-widget elementor-widget-divider\" data-id=\"ff24a28\" data-element_type=\"widget\" data-widget_type=\"divider.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t<style>\/*! elementor - v3.7.8 - 02-10-2022 *\/\n.elementor-widget-divider{--divider-border-style:none;--divider-border-width:1px;--divider-color:#2c2c2c;--divider-icon-size:20px;--divider-element-spacing:10px;--divider-pattern-height:24px;--divider-pattern-size:20px;--divider-pattern-url:none;--divider-pattern-repeat:repeat-x}.elementor-widget-divider .elementor-divider{display:-webkit-box;display:-ms-flexbox;display:flex}.elementor-widget-divider .elementor-divider__text{font-size:15px;line-height:1;max-width:95%}.elementor-widget-divider .elementor-divider__element{margin:0 var(--divider-element-spacing);-ms-flex-negative:0;flex-shrink:0}.elementor-widget-divider .elementor-icon{font-size:var(--divider-icon-size)}.elementor-widget-divider .elementor-divider-separator{display:-webkit-box;display:-ms-flexbox;display:flex;margin:0;direction:ltr}.elementor-widget-divider--view-line_icon .elementor-divider-separator,.elementor-widget-divider--view-line_text .elementor-divider-separator{-webkit-box-align:center;-ms-flex-align:center;align-items:center}.elementor-widget-divider--view-line_icon .elementor-divider-separator:after,.elementor-widget-divider--view-line_icon .elementor-divider-separator:before,.elementor-widget-divider--view-line_text .elementor-divider-separator:after,.elementor-widget-divider--view-line_text .elementor-divider-separator:before{display:block;content:\"\";border-bottom:0;-webkit-box-flex:1;-ms-flex-positive:1;flex-grow:1;border-top:var(--divider-border-width) var(--divider-border-style) var(--divider-color)}.elementor-widget-divider--element-align-left .elementor-divider .elementor-divider-separator>.elementor-divider__svg:first-of-type{-webkit-box-flex:0;-ms-flex-positive:0;flex-grow:0;-ms-flex-negative:100;flex-shrink:100}.elementor-widget-divider--element-align-left .elementor-divider-separator:before{content:none}.elementor-widget-divider--element-align-left .elementor-divider__element{margin-left:0}.elementor-widget-divider--element-align-right .elementor-divider .elementor-divider-separator>.elementor-divider__svg:last-of-type{-webkit-box-flex:0;-ms-flex-positive:0;flex-grow:0;-ms-flex-negative:100;flex-shrink:100}.elementor-widget-divider--element-align-right .elementor-divider-separator:after{content:none}.elementor-widget-divider--element-align-right .elementor-divider__element{margin-right:0}.elementor-widget-divider:not(.elementor-widget-divider--view-line_text):not(.elementor-widget-divider--view-line_icon) .elementor-divider-separator{border-top:var(--divider-border-width) var(--divider-border-style) var(--divider-color)}.elementor-widget-divider--separator-type-pattern{--divider-border-style:none}.elementor-widget-divider--separator-type-pattern.elementor-widget-divider--view-line .elementor-divider-separator,.elementor-widget-divider--separator-type-pattern:not(.elementor-widget-divider--view-line) .elementor-divider-separator:after,.elementor-widget-divider--separator-type-pattern:not(.elementor-widget-divider--view-line) .elementor-divider-separator:before,.elementor-widget-divider--separator-type-pattern:not([class*=elementor-widget-divider--view]) .elementor-divider-separator{width:100%;min-height:var(--divider-pattern-height);-webkit-mask-size:var(--divider-pattern-size) 100%;mask-size:var(--divider-pattern-size) 100%;-webkit-mask-repeat:var(--divider-pattern-repeat);mask-repeat:var(--divider-pattern-repeat);background-color:var(--divider-color);-webkit-mask-image:var(--divider-pattern-url);mask-image:var(--divider-pattern-url)}.elementor-widget-divider--no-spacing{--divider-pattern-size:auto}.elementor-widget-divider--bg-round{--divider-pattern-repeat:round}.rtl .elementor-widget-divider .elementor-divider__text{direction:rtl}.e-container>.elementor-widget-divider{width:var(--container-widget-width,100%);--flex-grow:var(--container-widget-flex-grow,0)}<\/style>\t\t<div class=\"elementor-divider\">\n\t\t\t<span class=\"elementor-divider-separator\">\n\t\t\t\t\t\t<\/span>\n\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-f731001 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"f731001\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-d1d52ca\" data-id=\"d1d52ca\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-d7ca55d elementor-widget elementor-widget-text-editor\" data-id=\"d7ca55d\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>CARACTERISTICAS<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-dcc2241 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"dcc2241\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-fd05c64\" data-id=\"fd05c64\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-09f5b78 elementor-widget elementor-widget-text-editor\" data-id=\"09f5b78\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>Descripci\u00f3n general de los aspectos m\u00e1s importantes de la etiqueta de afinidad de prote\u00ednas<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-733421f elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"733421f\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-407f33f\" data-id=\"407f33f\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-f852ecc elementor-widget elementor-widget-text-editor\" data-id=\"f852ecc\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<table class=\"responsiv mb-5\"><tbody><tr><th colspan=\"4\">CARACTERISTICAS DE HIS-TAG:<\/th><\/tr><tr><td>Amino Acid Sequence<\/td><td>HHHHHH (4-10)<\/td><\/tr><tr><td>DNA Sequence<\/td><td>5&#8242;-CAT CAC CAT CAC CAT CAC-3&#8242;<\/td><\/tr><tr><td>Size<\/td><td>840.8 Da (His6)<\/td><\/tr><tr><td>Compatibility to recombinant proteins<\/td><td>Can be added either to the N- or C-Terminus of a protein<\/td><\/tr><tr><td>Specificity of interaction (KD)<\/td><td>10 \u00b5M<\/td><\/tr><tr><td>Protein yield per ml of high-quality purification resin<\/td><td>Up to 80 mg \/ ml (GFP-His)<\/td><\/tr><tr><td>Elution conditions<\/td><td>Imidazole, histidine, or pH shift (pH 4-5)<\/td><\/tr><\/tbody><\/table>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-ef6d4c8 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"ef6d4c8\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-1aab683\" data-id=\"1aab683\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-a73954b elementor-widget elementor-widget-spacer\" data-id=\"a73954b\" data-element_type=\"widget\" data-widget_type=\"spacer.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t<div class=\"elementor-spacer\">\n\t\t\t<div class=\"elementor-spacer-inner\"><\/div>\n\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-c786a15 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"c786a15\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-3652195\" data-id=\"3652195\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-414e178 elementor-widget elementor-widget-text-editor\" data-id=\"414e178\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>\u00bfC\u00f3mo funciona una etiqueta His?<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-ca217fd elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"ca217fd\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-ec8de9a\" data-id=\"ec8de9a\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-d0eba41 elementor-widget elementor-widget-text-editor\" data-id=\"d0eba41\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>La purificaci\u00f3n de una prote\u00edna a trav\u00e9s del marcado con his se logra a trav\u00e9s de una interacci\u00f3n entre varios iones met\u00e1licos divalentes, como Ni2+ (el ion predominantemente utilizado) y el anillo b\u00e1sico de imidazol de la histidina. Las etiquetas His pueden unirse a esos metales a un pH neutro o ligeramente b\u00e1sico (7.5 &#8211; 8 es lo t\u00edpico). Vale la pena se\u00f1alar que la afinidad de uni\u00f3n entre esos dos socios de interacci\u00f3n es proporcional a la longitud de la etiqueta His. Se logra una afinidad de uni\u00f3n diez veces mayor con las etiquetas His10 en comparaci\u00f3n con las etiquetas His6 (Guignet, Hovius &amp; Vogel, 2004; Fessenden, 2009). No obstante, como primera prueba, se recomienda minimizar el n\u00famero de residuos de histidina para evitar posibles perturbaciones en la funci\u00f3n de la prote\u00edna (Bornhorst &amp; Falke, 2000).<\/p><p>Durante las \u00faltimas etapas del proceso de purificaci\u00f3n, la prote\u00edna se puede eluir disminuyendo el pH a 4-5, aunque cabe se\u00f1alar que algunas prote\u00ednas se desnaturalizan a un pH tan bajo. Adem\u00e1s, la mayor\u00eda de los cient\u00edficos prefieren una eluci\u00f3n por competici\u00f3n con concentraciones m\u00e1s altas de imidazol (100-500 mM). Tambi\u00e9n se recomienda insertar molaridades bajas de imidazol (5-20 mM, rara vez hasta 80 mM) en los tampones de carga y lavado para reducir los eventos de uni\u00f3n no espec\u00edfica, ya que algunas prote\u00ednas end\u00f3genas o histidinas naturales pueden mostrar una uni\u00f3n d\u00e9bil a IMAC. resinas (Bola\u00f1os-Garcia &amp; Davies, 2006). Las condiciones \u00f3ptimas para cada prote\u00edna siempre deben determinarse individualmente.<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-3ba0869 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"3ba0869\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-27bec98\" data-id=\"27bec98\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-b9c1711 elementor-widget elementor-widget-spacer\" data-id=\"b9c1711\" data-element_type=\"widget\" data-widget_type=\"spacer.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t<div class=\"elementor-spacer\">\n\t\t\t<div class=\"elementor-spacer-inner\"><\/div>\n\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-c647f3f elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"c647f3f\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-5f536e3\" data-id=\"5f536e3\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-630e4b0 elementor-widget elementor-widget-text-editor\" data-id=\"630e4b0\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>La mayor\u00eda de los protocolos de purificaci\u00f3n por afinidad siguen los mismos tres pasos (Fig. 2):<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-9b22ce6 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"9b22ce6\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-81f4c40\" data-id=\"81f4c40\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-5f681d6 elementor-widget elementor-widget-text-editor\" data-id=\"5f681d6\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<div class=\"row mt-4\"><div class=\"col-2 float-left text-center\">1. Vinculaci\u00f3n:<br \/>Se aplica a la columna una soluci\u00f3n cruda que contiene la prote\u00edna etiquetada con His y se une en funci\u00f3n de la interacci\u00f3n etiqueta de afinidad &#8211; matriz.<br \/>2. Lavado:<br \/>Otras prote\u00ednas que se unen de forma inespec\u00edfica se eliminan por lavado con tampones de lavado adecuados (ver prote\u00edna magenta en la Fig. 2). Estos tampones ya deber\u00edan contener molaridades bajas de imidazol.<br \/>3. Eluci\u00f3n:<br \/>La prote\u00edna unida espec\u00edficamente se eluye de la columna, normalmente mediante la uni\u00f3n competitiva de una mol\u00e9cula similar (p. ej., histidina e imidazol), cortando la etiqueta con una proteasa o mediante la desestabilizaci\u00f3n de la interacci\u00f3n etiqueta de afinidad &#8211; matriz, p. por un cambio de pH. El eluato puro se puede recolectar en tubos de reacci\u00f3n para futuras investigaciones.<\/div><\/div>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-b85a4c8 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"b85a4c8\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-0d08462\" data-id=\"0d08462\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-81fd39c elementor-widget elementor-widget-image\" data-id=\"81fd39c\" data-element_type=\"widget\" data-widget_type=\"image.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t\t\t\t\t\t<figure class=\"wp-caption\">\n\t\t\t\t\t\t\t\t\t\t<img width=\"600\" height=\"379\" src=\"https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/His-tag-Abbildung-2-600x379.jpg\" class=\"attachment-classic-grid-listing size-classic-grid-listing\" alt=\"\" loading=\"lazy\" srcset=\"https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/His-tag-Abbildung-2-600x379.jpg 600w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/His-tag-Abbildung-2-300x190.jpg 300w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/His-tag-Abbildung-2-1024x647.jpg 1024w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/His-tag-Abbildung-2-768x486.jpg 768w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/His-tag-Abbildung-2-253x160.jpg 253w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/His-tag-Abbildung-2.jpg 1112w\" sizes=\"(max-width: 600px) 100vw, 600px\" \/>\t\t\t\t\t\t\t\t\t\t\t<figcaption class=\"widget-image-caption wp-caption-text\">Figura 2: proceso Bind-Wash-Elute de una purificaci\u00f3n con etiqueta His. La concentraci\u00f3n de imidazol (naranja) aument\u00f3 con el tiempo para eluir la prote\u00edna de inter\u00e9s despu\u00e9s de realizar el lavado. Adem\u00e1s de la eluci\u00f3n de un paso, tambi\u00e9n es posible eluir la prote\u00edna con un gradiente. Esto es especialmente \u00fatil cuando se necesitan purezas elevadas.<\/figcaption>\n\t\t\t\t\t\t\t\t\t\t<\/figure>\n\t\t\t\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-32819e8 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"32819e8\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-692dd24\" data-id=\"692dd24\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-9facbd9 elementor-widget-divider--view-line elementor-widget elementor-widget-divider\" data-id=\"9facbd9\" data-element_type=\"widget\" data-widget_type=\"divider.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t<div class=\"elementor-divider\">\n\t\t\t<span class=\"elementor-divider-separator\">\n\t\t\t\t\t\t<\/span>\n\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-bf4da07 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"bf4da07\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-4e064ab\" data-id=\"4e064ab\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-2b655e3 elementor-widget elementor-widget-spacer\" data-id=\"2b655e3\" data-element_type=\"widget\" data-widget_type=\"spacer.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t<div class=\"elementor-spacer\">\n\t\t\t<div class=\"elementor-spacer-inner\"><\/div>\n\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-92401be elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"92401be\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-a79220f\" data-id=\"a79220f\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-cab4b35 elementor-widget elementor-widget-text-editor\" data-id=\"cab4b35\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>Purificaci\u00f3n de prote\u00ednas a trav\u00e9s de His-tag<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-4859f20 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"4859f20\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-de01c63\" data-id=\"de01c63\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-b2897c5 elementor-widget elementor-widget-text-editor\" data-id=\"b2897c5\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>\u00bfC\u00f3mo se purifica una prote\u00edna con una etiqueta His?<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-58ab2d3 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"58ab2d3\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-5430f2c\" data-id=\"5430f2c\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-ca3e60d elementor-widget elementor-widget-text-editor\" data-id=\"ca3e60d\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>La purificaci\u00f3n de prote\u00ednas a trav\u00e9s de His-tag a menudo se logra mediante una cromatograf\u00eda de afinidad de metal inmovilizado o, en resumen, IMAC. Este es un m\u00e9todo de purificaci\u00f3n ampliamente utilizado para separar prote\u00ednas y p\u00e9ptidos que muestran afinidad por los iones met\u00e1licos (como Ni2+ o Co2+) de una soluci\u00f3n. El marcado con His de una prote\u00edna permite que las prote\u00ednas de fusi\u00f3n reci\u00e9n formadas se unan a los metales quelados, lo que da como resultado enriquecimientos sustanciales en muy pocos pasos. Los quelantes comunes utilizados como ligandos son las resinas de agarosa IDA y NTA. Se pueden emparejar f\u00e1cilmente con un metal divalente, dependiendo de la relaci\u00f3n de afinidad\/especificidad deseada del proceso (ver Fig. 4).<br \/>Tabla 2: Comparaci\u00f3n entre tres ligandos y sus capacidades de uni\u00f3n al n\u00edquel<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-3f1fa31 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"3f1fa31\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-77455e6\" data-id=\"77455e6\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-b474cd3 elementor-widget elementor-widget-text-editor\" data-id=\"b474cd3\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<table class=\"responsiv\"><tbody><tr><th>LIGAND<\/th><th>COORDINATION SITES<\/th><th>NICKEL BINDING<\/th><\/tr><tr><td>Iminodiacetic acid (IDA)<\/td><td>3<\/td><td>Weak<\/td><\/tr><tr><td>Nitrilotriacetic acid (NTA)<\/td><td>4<\/td><td>Moderate<\/td><\/tr><tr><td>INDIGO-Ni<\/td><td>5<\/td><td>Strong<\/td><\/tr><\/tbody><\/table>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-4c9d956 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"4c9d956\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-4b28e29\" data-id=\"4b28e29\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-c6d5171 elementor-widget elementor-widget-spacer\" data-id=\"c6d5171\" data-element_type=\"widget\" data-widget_type=\"spacer.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t<div class=\"elementor-spacer\">\n\t\t\t<div class=\"elementor-spacer-inner\"><\/div>\n\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-246c162 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"246c162\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-6bb93b7\" data-id=\"6bb93b7\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-5ec9fa9 elementor-widget elementor-widget-text-editor\" data-id=\"5ec9fa9\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p><strong>\u2022 El ligando:<\/strong> esta es la mol\u00e9cula a la que se acopla el ion met\u00e1lico en un complejo quelante (ver Fig. 3).<br \/>Cube Biotech ofrece tres ligandos diferentes:<br \/>\u00a0 \u00a0 \u00a0 \u00a0 \u25e6 NTA: El \u00e1cido nitrilotriac\u00e9tico es el ligando m\u00e1s utilizado. Su complejo quelante con n\u00edquel se muestra en la Figura 3.<br \/>\u00a0 \u00a0 \u00a0 \u00a0 \u25e6 IDA: El \u00e1cido iminodiac\u00e9tico es el segundo m\u00e1s utilizado. Conozca las diferencias entre IDA y NTA aqu\u00ed.<br \/>\u00a0 \u00a0 \u00a0 \u00a0 \u25e6 INDIGO: Este ligando fue desarrollado aqu\u00ed en Cube Biotech. Entre sus ventajas se encuentran una tolerancia al DTT y al EDTA muy aumentada. Obtenga m\u00e1s informaci\u00f3n sobre este nuevo ligando.<\/p><p><strong>\u2022 El ion met\u00e1lico acoplado:<\/strong> seg\u00fan el ion met\u00e1lico acoplado al ligando, la afinidad y la especificidad de los cambios de purificaci\u00f3n de etiquetas His. El cobre tiene la afinidad m\u00e1s alta, mientras que a su vez tiene la especificidad m\u00e1s baja. El cobalto marca el otro extremo del espectro. El n\u00edquel, el i\u00f3n met\u00e1lico m\u00e1s utilizado, tiene un buen equilibrio entre afinidad y especificidad. Regla general: cuanto mayor sea el rendimiento de prote\u00edna, menos espec\u00edfica se vuelve la purificaci\u00f3n. La figura 4 puede brindarle una descripci\u00f3n general de la proporci\u00f3n de afinidad\/especificidad de los iones met\u00e1licos m\u00e1s utilizados para la purificaci\u00f3n de etiquetas His.<\/p><p><strong>\u2022 El tama\u00f1o de las perlas:<\/strong> cuanto mayor sea el di\u00e1metro de las perlas utilizadas, mayor ser\u00e1 el caudal para la purificaci\u00f3n por FPLC. Esto los hace m\u00e1s \u00fatiles para medios celulares viscosos o purificaci\u00f3n por lotes. Sin embargo, hacer uso de una velocidad de flujo m\u00e1s lenta generalmente aumenta el rendimiento de prote\u00edna, ya que las perlas m\u00e1s peque\u00f1as tienen una mayor relaci\u00f3n superficie-volumen. Por lo tanto, el mismo volumen de resina puede unir m\u00e1s prote\u00ednas que perlas de mayor tama\u00f1o. Ofrecemos di\u00e1metros de perlas de 30 \u00b5m, 40 \u00b5m, 100 \u00b5m y tama\u00f1o XL (~400 \u00b5m). Otros tama\u00f1os est\u00e1n disponibles bajo pedido.<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-e72141e elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"e72141e\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-c9afc30\" data-id=\"c9afc30\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-e974a82 elementor-widget-divider--view-line elementor-widget elementor-widget-divider\" data-id=\"e974a82\" data-element_type=\"widget\" data-widget_type=\"divider.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t<div class=\"elementor-divider\">\n\t\t\t<span class=\"elementor-divider-separator\">\n\t\t\t\t\t\t<\/span>\n\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-d3564d0 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"d3564d0\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-2e67c3f\" data-id=\"2e67c3f\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-b7c066f elementor-widget elementor-widget-text-editor\" data-id=\"b7c066f\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>Comparaci\u00f3n de la estructura del complejo quelante y los iones met\u00e1licos:<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-16f4a5b elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"16f4a5b\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-e7ca2d2\" data-id=\"e7ca2d2\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<section class=\"elementor-section elementor-inner-section elementor-element elementor-element-41159ce elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"41159ce\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-50 elementor-inner-column elementor-element elementor-element-e7d336e\" data-id=\"e7d336e\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-a347fd6 elementor-widget elementor-widget-image\" data-id=\"a347fd6\" data-element_type=\"widget\" data-widget_type=\"image.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t\t\t\t\t\t\t\t\t<img width=\"722\" height=\"520\" src=\"https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/His-tag-Abbildung-3.jpg\" class=\"attachment-large size-large\" alt=\"\" loading=\"lazy\" srcset=\"https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/His-tag-Abbildung-3.jpg 722w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/His-tag-Abbildung-3-300x216.jpg 300w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/His-tag-Abbildung-3-600x432.jpg 600w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/His-tag-Abbildung-3-222x160.jpg 222w\" sizes=\"(max-width: 722px) 100vw, 722px\" \/>\t\t\t\t\t\t\t\t\t\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t<div class=\"elementor-column elementor-col-50 elementor-inner-column elementor-element elementor-element-3843195\" data-id=\"3843195\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-0577241 elementor-widget elementor-widget-image\" data-id=\"0577241\" data-element_type=\"widget\" data-widget_type=\"image.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t\t\t\t\t\t<figure class=\"wp-caption\">\n\t\t\t\t\t\t\t\t\t\t<img width=\"800\" height=\"523\" src=\"https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/His-tag-Abbildung-4-1024x669.jpg\" class=\"attachment-large size-large\" alt=\"\" loading=\"lazy\" srcset=\"https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/His-tag-Abbildung-4-1024x669.jpg 1024w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/His-tag-Abbildung-4-300x196.jpg 300w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/His-tag-Abbildung-4-768x502.jpg 768w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/His-tag-Abbildung-4-600x392.jpg 600w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/His-tag-Abbildung-4-245x160.jpg 245w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/His-tag-Abbildung-4.jpg 1294w\" sizes=\"(max-width: 800px) 100vw, 800px\" \/>\t\t\t\t\t\t\t\t\t\t\t<figcaption class=\"widget-image-caption wp-caption-text\">Figura 3 (Estructura): Representaci\u00f3n esquem\u00e1tica de un complejo quelante que consta de NTA y Ni2+. Este ion de n\u00edquel se une a dos anillos de imidazol de una etiqueta His que se ha agregado a una prote\u00edna. Figura 4 (diagrama): descripci\u00f3n general de los iones met\u00e1licos m\u00e1s utilizados para la purificaci\u00f3n de prote\u00ednas con etiqueta His. El ion met\u00e1lico elegido es siempre un compromiso entre afinidad y especificidad.<\/figcaption>\n\t\t\t\t\t\t\t\t\t\t<\/figure>\n\t\t\t\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-3877959 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"3877959\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-07b3b5d\" data-id=\"07b3b5d\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-6711cee elementor-widget-divider--view-line elementor-widget elementor-widget-divider\" data-id=\"6711cee\" data-element_type=\"widget\" data-widget_type=\"divider.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t<div class=\"elementor-divider\">\n\t\t\t<span class=\"elementor-divider-separator\">\n\t\t\t\t\t\t<\/span>\n\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-7c223c6 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"7c223c6\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-1a03e9d\" data-id=\"1a03e9d\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-b60650c elementor-widget elementor-widget-text-editor\" data-id=\"b60650c\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>Como a\u00f1adir His-Tag<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-536045c elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"536045c\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-2955e69\" data-id=\"2955e69\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-11a0254 elementor-widget elementor-widget-text-editor\" data-id=\"11a0254\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>La adici\u00f3n de una etiqueta de afinidad de polihistidina a una prote\u00edna se puede lograr mediante varios m\u00e9todos. Su peque\u00f1o tama\u00f1o y carga les permite integrarse f\u00e1cilmente en los genes objetivo mediante PCR o mutag\u00e9nesis dirigida al sitio.<\/p><p>Para lograr esto, deber\u00e1 crear un cebador con codones de histidina repetitivos <strong>(CAC\/CAT)<\/strong> junto a un cod\u00f3n de INICIO o DETENCI\u00d3N. Adem\u00e1s, se agregan a este fragmento varias bases de un extremo de su prote\u00edna de inter\u00e9s. La etiqueta de afinidad de hexahistidina reci\u00e9n incorporada agregar\u00e1 aproximadamente 0,9 kDa a su peso molecular. Para evitar la interferencia con la actividad de la prote\u00edna provocada por el marcado, se insertan conectores entre la etiqueta y la prote\u00edna (ver Fig. 5). Se dividen en tres grupos generales: enlazadores flexibles, enlazadores r\u00edgidos y enlazadores escindibles in vivo (Chen, Zaro &amp; Shen, 2013). Los enlazadores comunes del grupo flexible son <strong>gly-gly-gly<\/strong> o <strong>gly-ser-gly<\/strong>. Se recomienda variar su longitud o dejarlos completamente fuera para lograr los mejores resultados para su prote\u00edna espec\u00edfica.<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-a7cb357 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"a7cb357\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-13bcf12\" data-id=\"13bcf12\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-62c1349 elementor-widget elementor-widget-image\" data-id=\"62c1349\" data-element_type=\"widget\" data-widget_type=\"image.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t\t\t\t\t\t<figure class=\"wp-caption\">\n\t\t\t\t\t\t\t\t\t\t<img width=\"800\" height=\"334\" src=\"https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/His-tag-Abbildung-5.jpg\" class=\"attachment-large size-large\" alt=\"\" loading=\"lazy\" srcset=\"https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/His-tag-Abbildung-5.jpg 971w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/His-tag-Abbildung-5-300x125.jpg 300w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/His-tag-Abbildung-5-768x320.jpg 768w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/His-tag-Abbildung-5-600x250.jpg 600w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/His-tag-Abbildung-5-280x117.jpg 280w\" sizes=\"(max-width: 800px) 100vw, 800px\" \/>\t\t\t\t\t\t\t\t\t\t\t<figcaption class=\"widget-image-caption wp-caption-text\">Figura 5: Construcci\u00f3n de una His-Tag a\u00f1adida corriente arriba o corriente abajo de la secuencia espec\u00edfica de prote\u00edna conectada por un enlazador.<\/figcaption>\n\t\t\t\t\t\t\t\t\t\t<\/figure>\n\t\t\t\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-dd11d10 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"dd11d10\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-031983e\" data-id=\"031983e\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-d518bb8 elementor-widget elementor-widget-spacer\" data-id=\"d518bb8\" data-element_type=\"widget\" data-widget_type=\"spacer.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t<div class=\"elementor-spacer\">\n\t\t\t<div class=\"elementor-spacer-inner\"><\/div>\n\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-33e4363 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"33e4363\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-e631d7d\" data-id=\"e631d7d\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<section class=\"elementor-section elementor-inner-section elementor-element elementor-element-ec91fa2 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"ec91fa2\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-50 elementor-inner-column elementor-element elementor-element-6e15b13\" data-id=\"6e15b13\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-8e5ecba elementor-widget elementor-widget-text-editor\" data-id=\"8e5ecba\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>\u00a0<\/p><p>1. Vector que contiene la secuencia de polihistidina N-terminal (naranja)\/sitio de clonaci\u00f3n m\u00faltiple (azul claro)\/esqueleto del vector (verde)<\/p><p>\u00a0<\/p><p>\u00a0<\/p><p>\u00a0<\/p><p>\u00a0<\/p><p>\u00a0<\/p><p>\u00a0<\/p><p>2. Adici\u00f3n de una secuencia espec\u00edfica de prote\u00edna (PSS) flanqueada por sitios de restricci\u00f3n<\/p><p>\u00a0<\/p><p>\u00a0<\/p><p>\u00a0<\/p><p>\u00a0<\/p><p>\u00a0<\/p><p>\u00a0<\/p><p>3. Clonaci\u00f3n de su fragmento de c\u00f3digo en el vector de fusi\u00f3n<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t<div class=\"elementor-column elementor-col-50 elementor-inner-column elementor-element elementor-element-122b666\" data-id=\"122b666\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-56217ce elementor-widget elementor-widget-image\" data-id=\"56217ce\" data-element_type=\"widget\" data-widget_type=\"image.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t\t\t\t\t\t<figure class=\"wp-caption\">\n\t\t\t\t\t\t\t\t\t\t<img width=\"436\" height=\"704\" src=\"https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/Abbildung-6a-copy.jpg\" class=\"attachment-large size-large\" alt=\"\" loading=\"lazy\" srcset=\"https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/Abbildung-6a-copy.jpg 436w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/Abbildung-6a-copy-186x300.jpg 186w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/Abbildung-6a-copy-372x600.jpg 372w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/Abbildung-6a-copy-99x160.jpg 99w\" sizes=\"(max-width: 436px) 100vw, 436px\" \/>\t\t\t\t\t\t\t\t\t\t\t<figcaption class=\"widget-image-caption wp-caption-text\">Figura 6: Integraci\u00f3n de PSS en un vector que contiene una secuencia de polihistidina<\/figcaption>\n\t\t\t\t\t\t\t\t\t\t<\/figure>\n\t\t\t\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-690df55 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"690df55\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-bbcf3cd\" data-id=\"bbcf3cd\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-ba2c962 elementor-widget-divider--view-line elementor-widget elementor-widget-divider\" data-id=\"ba2c962\" data-element_type=\"widget\" data-widget_type=\"divider.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t<div class=\"elementor-divider\">\n\t\t\t<span class=\"elementor-divider-separator\">\n\t\t\t\t\t\t<\/span>\n\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-3d99596 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"3d99596\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-295f0fe\" data-id=\"295f0fe\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-b1dfc94 elementor-widget elementor-widget-text-editor\" data-id=\"b1dfc94\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>Aplicaciones His-Tag<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-f1a70c9 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"f1a70c9\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-038feea\" data-id=\"038feea\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-d7211a3 elementor-widget elementor-widget-text-editor\" data-id=\"d7211a3\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>Adem\u00e1s de su conocido uso como ayuda para la purificaci\u00f3n de prote\u00ednas, las etiquetas His desempe\u00f1an un papel en muchas otras aplicaciones de la biotecnolog\u00eda. Ejemplos notables son los ensayos desplegables (Pulldown Assays), la asistencia en la formaci\u00f3n de cristales o los m\u00e9todos de marcaje inmunol\u00f3gico.<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-385d3ea elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"385d3ea\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-ed77f19\" data-id=\"ed77f19\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-42bbbee elementor-widget elementor-widget-text-editor\" data-id=\"42bbbee\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<ul><li style=\"list-style-type: none;\"><ul><li>Pulldown assays: affinity tags like His6 or GST paired with an immobilized ligand (e.g. Ni<sup>2+<\/sup>\u00a0agarose resin) can be used to specifically bind to a target protein complex in order to purify and identify it (Louche, Salcedo &amp; Bigot, 2017)<\/li><\/ul><\/li><\/ul><ul><li style=\"list-style-type: none;\"><ul><li>Crystal formation: an abundance of proteins can be crystallized with an intact His-tag without a negative impact on the tagged target protein (Carson\u00a0<em>et al.<\/em>, 2007). Smits\u00a0<em>et al.<\/em>\u00a0even described cases where the His-tag assisted in the formation of crystals in 2008 (Smits, Mueller &amp; Grieshaber, 2008)<\/li><\/ul><\/li><\/ul><ul><li style=\"list-style-type: none;\"><ul><li>Immune labeling: hexahistidine tags are utilized in ELISA assays, Western blot detection, flow cytometry, immunohistochemistry, or immune fluorescence<\/li><\/ul><\/li><\/ul><ul><li style=\"list-style-type: none;\"><ul><li>Serial His-tagging: single subunits of a large protein can be tagged respectively to act as a marker for activity states in a gel (Howarth, Chinnapen &amp; Gerrow\u00a0<em>et al.<\/em>, 2006)<\/li><\/ul><\/li><\/ul><ul><li style=\"list-style-type: none;\"><ul><li>SDS-PAGE staining: metal-based fluorescence dyes facilitate the detection of a His-tagged peptide in a gel (Zhao, Hellman &amp; Zhan\u00a0<em>et al.<\/em>, 2010)<\/li><\/ul><\/li><\/ul><ul><li style=\"list-style-type: none;\"><ul><li>SPR immobilization strategies: surface plasmon resonance, used to determine kinetics and thermodynamics of two binding molecules, offers multiple possible immobilization strategies for protein attachment via His-Tag (Fischer, Leech, &amp; Hubbard, 2011)<\/li><\/ul><\/li><\/ul>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-6f0495c elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"6f0495c\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-85f7d5b\" data-id=\"85f7d5b\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-f9d5d61 elementor-widget elementor-widget-text-editor\" data-id=\"f9d5d61\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>FAQ<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-47bb6a0 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"47bb6a0\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-97e9201\" data-id=\"97e9201\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-b8a56e3 elementor-widget elementor-widget-text-editor\" data-id=\"b8a56e3\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p><strong>\u00bfCu\u00e1nto tiempo debe durar un enlazador a la etiqueta His?<\/strong><br \/>La longitud del enlazador\/espaciador depende de la accesibilidad estructural de la prote\u00edna diana. Las longitudes de cadena m\u00e1s largas tienen una mayor movilidad y, por lo tanto, son cruciales para las etiquetas con impedimento est\u00e9rico (ver Fig. 7).<\/p><p><strong>\u00bfD\u00f3nde debo colocar la etiqueta His?<\/strong><br \/>La etiqueta His se puede agregar al extremo N o C. La ubicaci\u00f3n \u00f3ptima es espec\u00edfica de la prote\u00edna y depende de la estructura molecular, ya que algunos sitios podr\u00edan perturbar la accesibilidad de la etiqueta.<\/p><p><strong>\u00bfC\u00f3mo cortar una etiqueta His?<\/strong><br \/>La etiqueta de afinidad se puede eliminar mediante el uso de un sitio de escisi\u00f3n de proteasa insertado entre la etiqueta de polihistidina y la prote\u00edna (Nikolov, Hu &amp; Lin et al., 1992).<\/p><p><strong>\u00bfPor qu\u00e9 cortar la etiqueta His?<\/strong><br \/>La etiqueta His a veces se escinde para asegurar una actividad proteica intacta y una cristalizaci\u00f3n m\u00e1s f\u00e1cil, pero aqu\u00ed no se puede aplicar una regla general. Debido al tama\u00f1o y la carga relativamente peque\u00f1os, las etiquetas de His rara vez interfieren con la actividad de la prote\u00edna. Adem\u00e1s, incluso se han descrito casos de formaci\u00f3n de cristales asistida por etiquetas de His. Sin embargo, para aplicaciones y usos farmac\u00e9uticos, a menudo se requiere que las prote\u00ednas est\u00e9n en su estado nativo, raz\u00f3n por la cual la escisi\u00f3n de la etiqueta His es un procedimiento est\u00e1ndar all\u00ed.<\/p><p><strong>\u00bfC\u00f3mo eluir etiquetas His de perlas?<\/strong><\/p><p>La eluci\u00f3n de las perlas se puede lograr con la ayuda de mayores concentraciones de imidazol, histidina o con un cambio de pH (pH 4 &#8211; 5).<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-69e72f9 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"69e72f9\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-fe118c4\" data-id=\"fe118c4\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-e9243b9 elementor-widget elementor-widget-image\" data-id=\"e9243b9\" data-element_type=\"widget\" data-widget_type=\"image.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t\t\t\t\t\t<figure class=\"wp-caption\">\n\t\t\t\t\t\t\t\t\t\t<img width=\"796\" height=\"619\" src=\"https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/short1.jpg\" class=\"attachment-large size-large\" alt=\"\" loading=\"lazy\" srcset=\"https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/short1.jpg 796w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/short1-300x233.jpg 300w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/short1-768x597.jpg 768w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/short1-600x467.jpg 600w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/short1-206x160.jpg 206w\" sizes=\"(max-width: 796px) 100vw, 796px\" \/>\t\t\t\t\t\t\t\t\t\t\t<figcaption class=\"widget-image-caption wp-caption-text\">Figura 7: Diferencias entre las longitudes de los enlazadores<\/figcaption>\n\t\t\t\t\t\t\t\t\t\t<\/figure>\n\t\t\t\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-f34ba68 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"f34ba68\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-043ff7c\" data-id=\"043ff7c\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-5a2736a elementor-widget elementor-widget-spacer\" data-id=\"5a2736a\" data-element_type=\"widget\" data-widget_type=\"spacer.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t<div class=\"elementor-spacer\">\n\t\t\t<div class=\"elementor-spacer-inner\"><\/div>\n\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-c42567c elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"c42567c\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-a74e1b3\" data-id=\"a74e1b3\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-b153c56 elementor-widget elementor-widget-text-editor\" data-id=\"b153c56\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>Referencias<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-26c5e2d elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"26c5e2d\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-0273976\" data-id=\"0273976\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-df575e5 elementor-widget elementor-widget-text-editor\" data-id=\"df575e5\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<ol><li>Hochuli, E., Bannwarth, W., D\u00f6beli, H.\u00a0<em>et al.<\/em>\u00a0Genetic Approach to Facilitate Purification of Recombinant Proteins with a Novel Metal Chelate Adsorbent.\u00a0<em>Nat Biotechnol<\/em>\u00a0<strong>6<\/strong>, 1321\u20131325 (1988).<\/li><li>Guignet, E.G., Hovius, R., Vogel, H. Reversible site-selective labeling of\u00a0<a href=\"https:\/\/cube-biotech.com\/membrane-proteins-indispensable-gatekeepers\">membrane proteins<\/a>\u00a0in live cells.\u00a0<em>Nature Biotechnology<\/em>,\u00a0<strong>22<\/strong>\u00a0(4), pp. 440-444 (2004)<\/li><li>Fessenden, JD., F\u00f6rster resonance energy transfer measurements of ryanodine receptor type 1 structure using a novel site-specific labeling method.\u00a0<em>PLoS One<\/em>,\u00a0<strong>4<\/strong>\u00a0(10), p. e7338 (2009)<\/li><li>Bolanos-Garcia, V.M., Davies, O.R. Structural analysis and classification of native proteins from E. coli commonly co-purified by immobilised metal affinity chromatography.\u00a0<em>Biochim. Biophys. Acta<\/em>,\u00a0<strong>1760<\/strong>, pp. 1304-1313 (2006)<\/li><li>David, N.E., Gee, M., Andersen, B.\u00a0<em>et al. J Biol, Chem<\/em>\u00a0<strong>272<\/strong>:15553 (1997).<\/li><li>Skerra A. A general vector, pASK84, for cloning, bacterial production, and single-step purification of antibody Fab fragments.\u00a0<em>Gene<\/em>,\u00a0<strong>141<\/strong>(1), 79\u201384 (1994).<\/li><li>Petty, K J., Metal-chelate affinity chromatography.\u00a0<em>Current protocols in neuroscience<\/em>\u00a0vol. Chapter\u00a0<strong>5<\/strong>\u00a0(2001)<\/li><li>Carson, M.\u00a0<em>et al.<\/em>, His-tag impact on structure.\u00a0<em>Acta crystallographica.<\/em>\u00a0Section D, Biological crystallography vol.\u00a0<strong>63<\/strong>, Pt 3 (2007)<\/li><li>Smits, S.H., Mueller, A., Grieshaber, M.K., Schmitt, L. Coenzyme- and His-tag-induced crystallization of octopine dehydrogenase\u00a0<em>Acta Crystallogr.<\/em>\u00a0Sect. F Struct. Biol. Cryst. Commun.,\u00a0<strong>64<\/strong>, pp. 836-839 (2008)<\/li><li>Howarth, M., Chinnapen, DF., Gerrow, K.\u00a0<em>et al.<\/em>\u00a0A monovalent streptavidin with a single femtomolar biotin binding site.\u00a0<em>Nat Methods<\/em>\u00a0<strong>3<\/strong>, 267\u2013273 (2006)<\/li><li>Chen, X., Zaro, J.L., and Shen, W.-C. Fusion protein linkers: property, design and functionality\u00a0<em>Adv. Drug Delivery Rev.<\/em>\u00a0<strong>65<\/strong>, 1357\u2013 1369 (2013)<\/li><li>Bornhorst, J. A. &amp; Falke, J. J. Purification of proteins using polyhistidine affinity tags.\u00a0<em>Methods in enzymology<\/em>,\u00a0<strong>326<\/strong>, 245\u2013254 (2000)<\/li><li>Louche A., Salcedo S.P., Bigot S. Protein\u2013Protein Interactions: Pull-Down Assays.\u00a0<em>Bacterial Protein Secretion Systems. Methods in Molecular Biology<\/em>, vol\u00a0<strong>1615<\/strong>\u00a0(2017)<\/li><li>Zhao, C., Hellman, L.M., Zhan, X., Bowman, W.S., Whiteheart, S.W., Fried, M.G. Hexahistidine-tag-specific optical probes for analyses of proteins and their interactions.\u00a0<em>Analytical Biochemistry<\/em>, Vol.\u00a0<strong>399<\/strong>, Issue 2 (2010)<\/li><li>Fischer, M., Leech, A. P. &amp; Hubbard, R. E. Comparative assessment of different histidine-tags for immobilization of protein onto surface plasmon resonance sensorchips.\u00a0<em>Analytical chemistry<\/em>,\u00a0<strong>83<\/strong>(5), 1800\u20131807 (2011)<\/li><li>Nikolov, D. B., Hu, S. H., Lin, J.\u00a0<em>et al.<\/em>\u00a0Crystal structure of TFIID TATA-box binding protein.\u00a0<em>Nature<\/em>\u00a0<strong>360<\/strong>, 6399 (1992)<\/li><\/ol>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-b7beaee elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"b7beaee\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-4fc36bc\" data-id=\"4fc36bc\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-7300873 elementor-widget elementor-widget-spacer\" data-id=\"7300873\" data-element_type=\"widget\" data-widget_type=\"spacer.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t<div class=\"elementor-spacer\">\n\t\t\t<div class=\"elementor-spacer-inner\"><\/div>\n\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-7d2d689 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"7d2d689\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-c54dc5a\" data-id=\"c54dc5a\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-81029c6 elementor-widget elementor-widget-image\" data-id=\"81029c6\" data-element_type=\"widget\" data-widget_type=\"image.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t\t\t\t\t\t\t\t\t<img width=\"255\" height=\"41\" src=\"https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/logo-small.jpg\" class=\"attachment-large size-large\" alt=\"\" loading=\"lazy\" \/>\t\t\t\t\t\t\t\t\t\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t\t\t\t<\/div>\n\t\t","protected":false},"excerpt":{"rendered":"<p>La etiqueta de polihistidina o His6 es una etiqueta de prote\u00edna desarrollada originalmente para la purificaci\u00f3n eficiente de prote\u00ednas en 1988 (Hochuli, Bannwarth &amp; D\u00f6beli et al., 1988). Como resultado de su relaci\u00f3n costo-beneficio comparablemente grande, es conocida como una de las etiquetas de afinidad de prote\u00ednas m\u00e1s utilizadas hasta la fecha. Debido a su [&hellip;]<\/p>\n","protected":false},"author":2,"featured_media":0,"parent":0,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"om_disable_all_campaigns":false,"_monsterinsights_skip_tracking":false,"_monsterinsights_sitenote_active":false,"_monsterinsights_sitenote_note":"","_monsterinsights_sitenote_category":0},"aioseo_notices":[],"_links":{"self":[{"href":"https:\/\/nuevo22.cidsamexico.com\/index.php\/wp-json\/wp\/v2\/pages\/2499"}],"collection":[{"href":"https:\/\/nuevo22.cidsamexico.com\/index.php\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/nuevo22.cidsamexico.com\/index.php\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/nuevo22.cidsamexico.com\/index.php\/wp-json\/wp\/v2\/users\/2"}],"replies":[{"embeddable":true,"href":"https:\/\/nuevo22.cidsamexico.com\/index.php\/wp-json\/wp\/v2\/comments?post=2499"}],"version-history":[{"count":21,"href":"https:\/\/nuevo22.cidsamexico.com\/index.php\/wp-json\/wp\/v2\/pages\/2499\/revisions"}],"predecessor-version":[{"id":11037,"href":"https:\/\/nuevo22.cidsamexico.com\/index.php\/wp-json\/wp\/v2\/pages\/2499\/revisions\/11037"}],"wp:attachment":[{"href":"https:\/\/nuevo22.cidsamexico.com\/index.php\/wp-json\/wp\/v2\/media?parent=2499"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}