{"id":8641,"date":"2023-03-15T20:00:28","date_gmt":"2023-03-16T02:00:28","guid":{"rendered":"https:\/\/nuevo22.cidsamexico.com\/?post_type=al_product&p=8641"},"modified":"2023-03-15T21:39:08","modified_gmt":"2023-03-16T03:39:08","slug":"lisado-de-e-coli-libre-de-celulas","status":"publish","type":"al_product","link":"https:\/\/nuevo22.cidsamexico.com\/index.php\/productos\/lisado-de-e-coli-libre-de-celulas\/","title":{"rendered":"Lisado de E.coli libre de c\u00e9lulas\u00a0"},"content":{"rendered":"\n

Nuestros lisados libres de c\u00e9lulas de E.coli son la maquinaria de purificaci\u00f3n de prote\u00ednas aisladas de c\u00e9lulas bacterianas. Crean un sistema de expresi\u00f3n de prote\u00ednas libre de c\u00e9lulas ajustable y de alto rendimiento. Las ventajas de la expresi\u00f3n de prote\u00ednas libres de c\u00e9lulas son numerosas, entre otras cosas, permite a un cient\u00edfico: <\/p>\n\n\n\n

\u2022 Para la estabilizaci\u00f3n directa y la integraci\u00f3n cotraduccional de prote\u00ednas de membrana en nanodiscos.<\/p>\n\n\n\n

\u2022 Expresi\u00f3n de prote\u00ednas que de otro modo ser\u00edan t\u00f3xicas<\/p>\n\n\n\n

<\/div>\n\n\n\n
Catalogo<\/th>Producto<\/th>Presentaci\u00f3n<\/th><\/tr><\/thead>
CU-21001<\/td>Cell-free E.coli<\/em> lysate-45 <\/td>350 \u00b5l<\/td><\/tr>
CU-21011<\/td>Cell-free E.coli<\/em> lysate HighYield<\/td>350 \u00b5l<\/td><\/tr>
CU-21033<\/td>Cell-free E.coli<\/em> lysate HighYield-T7<\/td>4 x 330 \u00b5l<\/td><\/tr><\/tbody><\/table><\/figure>\n\n\n\n
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\"pdf\"\/<\/figure>\n\n\n\n

Protocol T7<\/a> <\/p>\n\n\n\n

\"pdf\"\/<\/figure>\n\n\n\n

Protocol<\/a><\/p>\n<\/div>\n\n\n\n

Datasheet a petici\u00f3n, ya que son espec\u00edficas del lote.<\/p>\n\n\n\n

<\/div>\n\n\n\n
<\/div>\n\n\n\n
FEATURE<\/th><\/th><\/tr>
State<\/td>Polvo liofilizado
para disolver en agua<\/td><\/tr>
Peso Molecular<\/td>10-11 kDa<\/td><\/tr>
Buffer<\/td>HEPES<\/td><\/tr>
pH after solving<\/td>7.5<\/td><\/tr>
Divalent cationic tolerance<\/td>6 mM<\/td><\/tr>
Acrylic Acid to Styrene Ratio<\/td>45% a 55%<\/td><\/tr>
Almacenamiento (periodo largo)<\/td>4\u00b0C <\/td><\/tr><\/tbody><\/table><\/figure>\n\n\n\n
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Estructura <\/figcaption><\/figure>\n\n\n\n
<\/div>\n\n\n\n

Resultado de Laboratotio<\/mark><\/p>\n\n\n\n

En estrecha colaboraci\u00f3n con el grupo de Volker Doetsch y Frank Bernhard, Instituto de Biof\u00edsica de la Universidad de Frankfurt, Alemania, proporcionamos un sistema abierto que se puede adaptar para una variedad de aplicaciones, con protocolos dedicados para prote\u00ednas de membrana, intercambio continuo (di\u00e1lisis ) reacciones, y muchos m\u00e1s. Este sistema requiere m\u00e1s trabajo inicialmente que los kits listos para usar, pero proporciona la m\u00e1xima flexibilidad en las variaciones de reacci\u00f3n.<\/p>\n\n\n\n

<\/div>\n\n\n\n
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\"\"<\/figure>\n\n\n\n

Figura 1.<\/strong> Expresi\u00f3n libre de c\u00e9lulas est\u00e1ndar frente a HiYield de GFP. 1,7: marcador de PM, 2-6: expresi\u00f3n de lisado de alto rendimiento; variaci\u00f3n de la concentraci\u00f3n de Mg2+ (14, 16, 18, 20,22 mM). 8-12: Expresi\u00f3n de lisado est\u00e1ndar, misma variaci\u00f3n de concentraciones de Mg2+. Se observ\u00f3 un \u00f3ptimo de Mg2+ a ca. 16 mM para estas combinaciones de lisados libres de c\u00e9lulas y otros componentes de reacci\u00f3n. Los rendimientos oscilan entre 1,3 y 5 mg\/ml para reacciones de alto rendimiento y entre 1 y 3,8 mg\/ml para reacciones est\u00e1ndar. Datos proporcionados amablemente por Frank Bernhard, Universidad de Frankfurt, Departamento de Qu\u00edmica Biof\u00edsica.<\/p>\n<\/div><\/div>\n\n\n\n

<\/div>\n\n\n\n

Equipo y Materiales requeridos adicinalmente<\/p>\n\n\n\n

Required equipment<\/td>Micropipettor and Micropipetting tips
Thermomix \/ shaking incubator for 500 rpm and 30\u00b0C incubation
Ultrasonic device (Optional)
Dialysis chambers and feeding mix reservoirs, depending on reaction scales. Alternatively home-built dialysis chambers combined with dialysis membrane, cutoff 12-14 kDa.<\/td><\/tr>
Additional materials & chemicals<\/td>1,4-Dithiotreitol (DTT)
Acetyl phosphate lithium potassium salt (ACP)
Acetic AcidAdenosine 5\u2019-triphosphate (ATP)
Amino acids for cell-free expression
Complete protease inhibitor cocktailCytidine 5\u2019-triphosphate di-sodium salt (CTP)
Dipotassium hydrogen phosphate (K2<\/sub>HPO4<\/sub>)
Ethylenediaminetetraacetic acid (EDTA)
Folinic acid calcium salt
Guanosine 5\u2019-triphosphate di-sodium salt (GTP)
2-(4-(2-Hydroxyethyl)- 1-piperazinyl)-ethane sulfonic acid (HEPES)
Magnesium acetate tetrahydrate (Mg(oAc)2<\/sub>)
PEG 8,000
Phosphoenol pyruvic acid monopotassium salt (PEP)
Potassium acetate (KoAc)
Potassium chloride (KCl)
Potassium dihydrogenphosphate (KH2<\/sub>PO4<\/sub>)
Potassium hydroxide (KOH)
Pyruvate kinase (PK)
RiboLock RNase inhibitor
Sodium azide (NaN3<\/sub>)
Sodium chloride
Tris base
tRNA E.coli<\/em> MRE 600
Uridine 5\u2019-triphosphate tri-sodium salt (UTP)<\/td><\/tr>
Project specific materials<\/td>Template DNA
Optional:<\/em> Positive control template DNA
Recommended:<\/em> Suitable affinity chromatography matrix or magnetic beads SDS-PAGE\/ Western Blot equipment and antibody
For membrane protein expressions: <\/em>Pre-assembled MSP nanodisc<\/a>
For membrane protein expressions: <\/em>Detergents, e.g. Decylmaltoside, dodecylmaltoside<\/a><\/td><\/tr><\/tbody><\/table><\/figure>\n\n\n\n
<\/div>\n\n\n\n

Bibligraf\u00eda<\/mark><\/p>\n\n\n\n

TITLE<\/strong><\/th>YEAR<\/strong><\/th>AUTHOR<\/strong><\/th><\/tr>
High-Level Cell-Free Production of Membrane Proteins with Nanodiscs<\/td>2013<\/td>Roos C., Kai L., Haberstock S., Proverbio D., Ghoshdastider U., Ma Y., Filipek S., Wang X., D\u00f6tsch V., Bernhard F.<\/a><\/td><\/tr>
Structural determinants of lipid specificity within Ups\/PRELI lipid transfer proteins<\/td>2019<\/td>Miliara X., Tatsuta T., Berry J-L., Rouse S.L.,Solak K., Chorev D.S., Wu D., Robinson C.V., Matthews S.<\/a><\/td><\/tr>
Systematic optimization of cell-free synthesized human endothelin B receptor folding<\/td>2019<\/td>Rues R-B., Dong F., D\u00f6tsch V., & Bernhard F.<\/a><\/td><\/tr>
Systematic optimization of cell-free synthesized human endothelin B receptor folding<\/td>2019<\/td>MacVicar T., Ohba Y., Nolte H., Mayer F.-C., Tatsuta T., Sprenger H.-G., Lindner B., Zhao Y., Li J., Bruns C., Kr\u00fcger M., Habich M., Rlemer J., Schwarzer R., Pasparakis M., Henscke S., Br\u00fcning J., Zamboni N., Langer T.<\/a><\/td><\/tr>
Chemical Blockage of the Mitochondrial Rhomboid Protease PARL by Novel Ketoamide Inhibitors Reveals Its Role in PINK1\/Parkin-Dependent Mitophagy<\/td>2022<\/td>Pol\u00e1chov\u00e1 E., Back K., Heuten E., Stanchev S., Tich\u00e1 A., Lampe P., Majer P., Langer T., Lemberg M.K., St\u0159\u00ed\u0161ovsk\u00fd K.<\/a><\/td><\/tr><\/tbody><\/table><\/figure>\n\n\n\n
<\/div>\n","protected":false},"excerpt":{"rendered":"

Cell-free E.coli lysate<\/p>\n","protected":false},"featured_media":8818,"template":"","al_product-cat":[39,34],"aioseo_notices":[],"_links":{"self":[{"href":"https:\/\/nuevo22.cidsamexico.com\/index.php\/wp-json\/wp\/v2\/al_product\/8641"}],"collection":[{"href":"https:\/\/nuevo22.cidsamexico.com\/index.php\/wp-json\/wp\/v2\/al_product"}],"about":[{"href":"https:\/\/nuevo22.cidsamexico.com\/index.php\/wp-json\/wp\/v2\/types\/al_product"}],"version-history":[{"count":5,"href":"https:\/\/nuevo22.cidsamexico.com\/index.php\/wp-json\/wp\/v2\/al_product\/8641\/revisions"}],"predecessor-version":[{"id":8828,"href":"https:\/\/nuevo22.cidsamexico.com\/index.php\/wp-json\/wp\/v2\/al_product\/8641\/revisions\/8828"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/nuevo22.cidsamexico.com\/index.php\/wp-json\/wp\/v2\/media\/8818"}],"wp:attachment":[{"href":"https:\/\/nuevo22.cidsamexico.com\/index.php\/wp-json\/wp\/v2\/media?parent=8641"}],"wp:term":[{"taxonomy":"al_product-cat","embeddable":true,"href":"https:\/\/nuevo22.cidsamexico.com\/index.php\/wp-json\/wp\/v2\/al_product-cat?post=8641"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}