{"id":3206,"date":"2023-01-11T11:25:31","date_gmt":"2023-01-11T17:25:31","guid":{"rendered":"https:\/\/nuevo22.cidsamexico.com\/?post_type=al_product&p=3206"},"modified":"2023-01-20T13:51:12","modified_gmt":"2023-01-20T19:51:12","slug":"amplite-fluorimetric-proteasome-20s-activity-assay-kit-green-fluorescence","status":"publish","type":"al_product","link":"https:\/\/nuevo22.cidsamexico.com\/index.php\/productos\/amplite-fluorimetric-proteasome-20s-activity-assay-kit-green-fluorescence\/","title":{"rendered":"Amplite\u00ae Fluorimetric Proteasome 20S Activity Assay Kit *Green Fluorescence*"},"content":{"rendered":"\n

Nuestro kit de ensayo de Proteasoma Fluorometric 20S es un ensayo fluorescente homog\u00e9neo que mide la actividad de la proteasa similar a la quimotripsina asociada con el complejo de proteasoma en c\u00e9lulas cultivadas o lisados \u200b\u200bcelulares. Este kit utiliza LLVY-R110 como indicador fluorog\u00e9nico de la actividad del proteasoma. La escisi\u00f3n de LLVY-R110 por el proteasoma genera el R110 fluorescente de color verde fuerte que se monitorea fluorom\u00e9tricamente a 520-530 nm con una excitaci\u00f3n de 480-500 nm. El kit proporciona todos los componentes esenciales con un protocolo de ensayo optimizado. <\/p>\n\n\n\n

El ensayo es robusto y se puede adaptar f\u00e1cilmente para ensayos de alto rendimiento para la evaluaci\u00f3n de la actividad del proteasoma o la detecci\u00f3n de inhibidores en c\u00e9lulas cultivadas o en soluci\u00f3n. <\/p>\n\n\n\n

El ensayo se puede realizar en un conveniente formato de placa de microtitulaci\u00f3n de fluorescencia de 96 y 384 pozos. La funci\u00f3n principal del proteasoma es degradar prote\u00ednas innecesarias o da\u00f1adas mediante prote\u00f3lisis, una reacci\u00f3n qu\u00edmica que rompe los enlaces pept\u00eddicos. La v\u00eda de degradaci\u00f3n proteos\u00f3mica es esencial para muchos procesos celulares, incluido el ciclo celular, la regulaci\u00f3n de la expresi\u00f3n g\u00e9nica y las respuestas al estr\u00e9s oxidativo. La forma m\u00e1s com\u00fan del proteasoma en esta v\u00eda es el proteasoma 26S, un complejo proteol\u00edtico dependiente de ATP, que contiene una estructura de part\u00edculas central 20S (700 kDa) y dos tapas reguladoras 19S (700 kDa). El n\u00facleo 20S contiene tres actividades proteol\u00edticas principales, incluidas las similares a la quimotripsina, la tripsina y la caspasa. Es responsable de la descomposici\u00f3n de prote\u00ednas clave involucradas en la apoptosis, la reparaci\u00f3n del ADN, la endocitosis y el control del ciclo celular.<\/p>\n\n\n\n

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Catalogo<\/th>Producto<\/th>Presentaci\u00f3n<\/th><\/tr><\/thead>
AAT-13456<\/td>Amplite\u00ae Fluorimetric Proteasome 20S Activity Assay Kit *Green Fluorescence*<\/td>100 pruebas<\/td><\/tr><\/tbody><\/table><\/figure>\n\n\n\n
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SDS<\/a><\/p>\n\n\n\n

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

Importante: Solo para uso en investigaci\u00f3n (RUO). <\/p>\n\n\n\n

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Plataforma<\/mark><\/p>\n\n\n\n

Lector de Microplacas de Fluorescencia<\/mark> <\/p>\n\n\n\n

Excitaci\u00f3n<\/td>490 nm<\/td><\/tr>
Emisi\u00f3n<\/td>525 nm<\/td><\/tr>
Cutoff<\/td>515 nm<\/td><\/tr>
Placa recomendada<\/td>S\u00f3lida negra <\/td><\/tr>
Especificaciones instrumento<\/td>Modo de lectura superior<\/td><\/tr><\/tbody><\/table><\/figure>\n\n\n\n
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Componentes<\/mark><\/p>\n\n\n\n

Componente A: Proteasome LLVY-R110 Substrate<\/td>1 vial <\/td><\/tr>
Componente B: Buffer de ensayo <\/td>1 botella (10 mL)<\/td><\/tr>
Componente C: DMSO<\/td>1 vial (100 \u00b5L)<\/td><\/tr><\/tbody><\/table><\/figure>\n\n\n\n
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Espectro<\/mark><\/p>\n\n\n\n

Abrir en Advanced Spectrum Viewer<\/a><\/em><\/p>\n\n\n\n

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Propiedades espectrales<\/mark><\/p>\n\n\n\n

Coeficiente de extinci\u00f3n (cm -1<\/sup> M -1<\/sup>)<\/td>80000<\/td><\/tr>
Excitaci\u00f3n (nm)<\/td>500<\/td><\/tr>
Emisi\u00f3n (nm)<\/td>522<\/td><\/tr><\/tbody><\/table><\/figure>\n\n\n\n
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PREPARACION DE SOLUCION DE STOCK<\/p>\n\n\n\n

A menos que se indique lo contrario, todas las soluciones madre no utilizadas deben dividirse en al\u00edcuotas de un solo uso y almacenarse a -20 \u00b0C despu\u00e9s de la preparaci\u00f3n. Evite los ciclos repetidos de congelaci\u00f3n y descongelaci\u00f3n.<\/em><\/p>\n\n\n\n

Proteasoma LLVY-R110 Soluci\u00f3n madre de sustrato (400X)<\/strong>
Agregue 25 \u00b5L de DMSO (Componente C) al vial de Proteasome LLVY-R110 Substrate (Component A) y mezcle bien para preparar una soluci\u00f3n madre de Proteasome LLVY-R110 Substrate 400X.<\/p>\n\n\n\n

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PREPARACION CELULAR<\/p>\n\n\n\n

Para obtener guia sobre la preparaci\u00f3n de muestras de c\u00e9lulas, visite: https:\/\/www.aatbio.com\/resources\/guides\/cell-sample-preparation.html<\/a><\/p>\n\n\n\n

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PREPARACION DE SOLUCION DE TRABAJO<\/p>\n\n\n\n

Agregue 25 \u03bcL de soluci\u00f3n madre de sustrato de Proteasome LLVY-R110 400X en 10 mL de buffer de ensayo (componente B) y mezcle bien para obtener una soluci\u00f3n de trabajo de Proteasome. Nota: Esta soluci\u00f3n de trabajo de Proteasoma es suficiente para 1 placa. Proteger de la luz.<\/p>\n\n\n\n

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Imagen<\/mark><\/p>\n\n\n\n

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Figura 1<\/strong>. Detecci\u00f3n de actividad de proteasoma en c\u00e9lulas Jurkat con el kit de ensayo de actividad de proteasoma 20S fluorim\u00e9trico Amplite\u00ae. Las c\u00e9lulas Jurkat se sembraron el mismo d\u00eda a raz\u00f3n de 500 000 c\u00e9lulas\/90 \u00b5l\/pocillo en una placa Costar de 96 pocillos de pared negra\/fondo transparente. Las c\u00e9lulas se trataron con o sin H2O2 50 mM durante 30 minutos. Se a\u00f1adi\u00f3 la soluci\u00f3n de carga del ensayo de proteasoma (100 \u00b5l\/pocillo) y se incub\u00f3 en incubadora de CO2 a 37 \u00b0C al 5 % durante 3 horas. La intensidad de la fluorescencia se midi\u00f3 a Ex\/Em = 490\/525 utilizando un lector de microplacas fluorescentes Gemini (Molecular Devices).<\/p>\n\n\n\n

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

Quantitative reactive cysteinome profiling reveals a functional link between ferroptosis and proteasome-mediated degradation<\/a>
Authors: <\/strong>Wang, Yankun and Wang, Chu
Journal: <\/strong>Cell Death \\& Differentiation (2022): 1–12<\/p>\n\n\n\n

TRIB2 modulates proteasome function to reduce ubiquitin stability and protect liver cancer cells against oxidative stress<\/a>
Authors: <\/strong>Guo, Susu and Chen, Yuxin and Yang, Yueyue and Zhang, Xiao and Ma, Lifang and Xue, Xiangfei and Qiao, Yongxia and Wang, Jiayi
Journal: <\/strong>Cell death \\& disease (2021): 1–18<\/p>\n\n\n\n

TRIB2 desensitizes ferroptosis via $\\beta$TrCP-mediated TFRC ubiquitiantion in liver cancer cells<\/a>
Authors: <\/strong>Guo, Susu and Chen, Yuxin and Xue, Xiangfei and Yang, Yueyue and Wang, Yikun and Qiu, Shiyu and Cui, Jiangtao and Zhang, Xiao and Ma, Lifang and Qiao, Yongxia and others,
Journal: <\/strong>Cell death discovery (2021): 1–13<\/p>\n\n\n\n

MnFe2O4 nanoparticles accelerate the clearance of mutant huntingtin selectively through ubiquitin-proteasome system<\/a>
Authors: <\/strong>Zhang, Li and Wei, Peng-Fei and Song, Yong-Hong and Dong, Liang and Wu, Ya-Dong and Hao, Zong-Yao and Fan, Song and Tai, Sheng and Meng, Jia-Lin and Lu, Yang and others, undefined
Journal: <\/strong>Biomaterials (2019): 119248<\/p>\n\n\n\n

Melatonin Promotes Ubiquitination of Phosphorylated Pro-Apoptotic Protein Bcl-2-Interacting Mediator of Cell Death-Extra Long (BimEL) in Porcine Granulosa Cells<\/a>
Authors: <\/strong>Wang, Yingzheng and Zeng, Shenming
Journal: <\/strong>International journal of molecular sciences (2018): 3431<\/p>\n\n\n\n

Photothermally Controlled MHC Class I Restricted CD8+ T-Cell Responses Elicited by Hyaluronic Acid Decorated Gold Nanoparticles as a Vaccine for Cancer Immunotherapy<\/a>
Authors: <\/strong>Cao, Fengqiang and Yan, Mengmeng and Liu, Yijia and Liu, Lanxia and Ma, Guilei
Journal: <\/strong>Advanced healthcare materials (2018): 1701439<\/p>\n\n\n\n

Mycobacterium tuberculosis controls phagosomal acidification by targeting CISH-mediated signaling<\/a>
Authors: <\/strong>Queval, Christophe J and Song, Ok-Ryul and Carralot, Jean-Philippe and Saliou, Jean-Michel and Bongiovanni, Antonino and Deloison, Gaspard and Deboosere, Nathalie and Jouny, Samuel and Iantomasi, Raffaella and Delorme, Vincent and others,
Journal: <\/strong>Cell reports (2017): 3188–3198<\/p>\n\n\n\n

Interactome analysis reveals a novel role for RAD6 in the regulation of proteasome activity and localization in response to DNA damage
Authors: <\/strong>An, Hongli and Yang, Lu and Wang, Chen and Gan, Zhixue and Gu, Haihui and Zhang, Tao and Huang, Xin and Liu, Yan and Li, Yufeng and Chang, Shing-Jyh and others,
Journal: <\/strong>Molecular and cellular biology (2017): e00419–16<\/p>\n\n\n\n

Mycobacterium tuberculosis Controls Phagosomal Acidification by Targeting CISH-Mediated Signaling<\/a>
Authors: <\/strong>Queval, Christophe J and Song, Ok-Ryul and Carralot, Jean-Philippe and Saliou, Jean-Michel and Bongiovanni, Antonino and Deloison, Gaspard and Deboos\u00e8re, Nathalie and Jouny, Samuel and Iantomasi, Raffaella and Delorme, Vincent and others, undefined
Journal: <\/strong>Cell Reports (2017): 3188–3198<\/p>\n\n\n\n

Autophagy-mediated clearance of ubiquitinated mutant huntingtin by graphene oxide<\/a>
Authors: <\/strong>Jin, Peipei and Wei, Pengfei and Zhang, Yunjiao and Lin, Jun and Sha, Rui and Hu, Yi and Zhang, Jiqian and Zhou, Wei and Yao, Han and Ren, Li and others, undefined
Journal: <\/strong>Nanoscale (2016)<\/p>\n\n\n\n

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Application Notes (en Ingles)<\/mark><\/p>\n\n\n\n

Interactome analysis reveals a n 1 ovel role for RAD6 in the regulation of proteasome activity and localization in response to DNA damage<\/a>
L-BMAA Induced ER Stress and Enhanced Caspase 12 Cleavage at Low Nonexcitotoxic Concentrations<\/a>
Interactome analysis reveals a n 1 ovel role for RAD6 in the regulation of proteasome activity and localization in response to DNA damage<\/a>
L-BMAA Induced ER Stress and Enhanced Caspase 12 Cleavage at Low Nonexcitotoxic Concentrations<\/a>
Interactome analysis reveals a n 1 ovel role for RAD6 in the regulation of proteasome activity and localization in response to DNA damage<\/a><\/p>\n\n\n\n

<\/div>\n","protected":false},"excerpt":{"rendered":"

Kit de ensayo para la actividad del proteasoma Proteasome 20S Activity Fluorometrico Amplite\u00ae *Fluorescencia verde* Nuestro kit es un ensayo fluorescente homog\u00e9neo que mide la actividad de la proteasa similar a la quimotripsina asociada con el complejo de proteasoma en c\u00e9lulas cultivadas o lisados \u200b\u200bcelulares. <\/p>\n","protected":false},"featured_media":3208,"template":"","al_product-cat":[34],"aioseo_notices":[],"_links":{"self":[{"href":"https:\/\/nuevo22.cidsamexico.com\/index.php\/wp-json\/wp\/v2\/al_product\/3206"}],"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":4,"href":"https:\/\/nuevo22.cidsamexico.com\/index.php\/wp-json\/wp\/v2\/al_product\/3206\/revisions"}],"predecessor-version":[{"id":3733,"href":"https:\/\/nuevo22.cidsamexico.com\/index.php\/wp-json\/wp\/v2\/al_product\/3206\/revisions\/3733"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/nuevo22.cidsamexico.com\/index.php\/wp-json\/wp\/v2\/media\/3208"}],"wp:attachment":[{"href":"https:\/\/nuevo22.cidsamexico.com\/index.php\/wp-json\/wp\/v2\/media?parent=3206"}],"wp:term":[{"taxonomy":"al_product-cat","embeddable":true,"href":"https:\/\/nuevo22.cidsamexico.com\/index.php\/wp-json\/wp\/v2\/al_product-cat?post=3206"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}