采用Surefire AlphaScreen® 试剂盒((TGR Biosciences and PerkinElmer)进行pERK检测

Cassandra Koole; Denise Wootten; Patrick Sexton

doi:10.21769/BioProtoc.806

Improve Research Reproducibility A Bio-protocol resource

提交稿件
订阅
登录
/
注册
- 个人主页
- 编辑个人信息
- 修改密码
- 退出
CN
- EN - English
- CN - 中文

Peer-reviewed

pERK Detection Assays Using the Surefire AlphaScreen^® Kit (TGR Biosciences and PerkinElmer)

采用Surefire AlphaScreen® 试剂盒((TGR Biosciences and PerkinElmer)进行pERK检测

CK Cassandra Koole email

DW Denise Wootten

PS Patrick Sexton

发布: 2013年07月05日第3卷第13期 DOI: 10.21769/BioProtoc.806 浏览次数: 10342

评审: Cheng Zhang

PDF

Q&A

引用

Cited by

参见作者原研究论文

The authors used this protocol in:

Cover of Proceedings of the National Academy of Sciences of the United States of America, featuring study using the protocol.

Nov 2012

实验方案合集

Cell Imaging - A Special Collection for Cell Bio 2023

相关实验方案

半透性人细胞中蛋白质和肽应激颗粒结合的定量测定

Saskia Hutten and Dorothee Dormann

2020年12月20日 4684 阅读

利用苯乙烯-马来酸脂质颗粒（SMALPs）提取富集膜蛋白用于下游分析

Benedict Dirnberger [...] Kathryn S. Lilley

2023年08月05日 1922 阅读

基于表面等离子体共振（SPR）生物传感器的人PD-1/PD-L1阻断检测方法的建立

Tess Puopolo [...] Chang Liu

2023年08月05日 2045 阅读

Abstract

Extracellular signal-regulated kinase 1 and 2 (ERK1/2) are serine/threonine protein kinases that are phosphorylated on Thr202/Tyr204 (ERK1) and Thr185/Tyr187 (ERK2). Phosphorylation of ERK1/2 (pERK1/2) arises from multiple stimuli, resulting in physiological responses that include cell growth, proliferation and differentiation. This protocol has been optimized for the detection of ligand-mediated pERK1/2 in adherent immortal cell lines expressing G protein-coupled receptors (GPCRs).

Materials and Reagents

Dulbecco’s modified eagle medium (DMEM) (Life Technologies, Gibco^®, catalog number: 11995-065 )
Bovine serum albumin (BSA) (Sigma-Aldrich, catalog number: A7906 )
SureFire^® Reagents (Includes Lysis, Activation and Reaction buffers) (TGR BioSciences, catalog number: TGRES500 )
AlphaScreen^® General IgG (Protein A) detection kit (PerkinElmer, catalog number: 6760617 )
White ProxiPlate 384-well microplate (PerkinElmer, catalog number: 6008280 )
TopSeal (PerkinElmer, catalog number: 6005250 )
NaCl
KCl
Na₂HPO₄
KH₂PO₄
Phosphate buffered saline (PBS) (see Recipes)
Detection buffer (see Recipes)

Equipment

Fusion-α plate reader or Envision plate reader with appropriate Alphascreen detection modules (PerkinElmer)
Sterile 96-well clear flat bottom plates (BD Biosciences, Falcon^®, catalog number: 353072 )
Humidified incubator
Multichannel pipettes
Micropipettes
Orbital shaker

Procedure

English

中文翻译

文章信息

版权信息

如何引用

Koole, C., Wootten, D. and Sexton, P. M. (2013). pERK Detection Assays Using the Surefire AlphaScreen^® Kit (TGR Biosciences and PerkinElmer). Bio-protocol 3(13): e806. DOI: 10.21769/BioProtoc.806.