論文

基本情報

氏名 竹谷 浩介
氏名(カナ) タケヤ コウスケ
氏名(英語) Takeya Kosuke
所属 獣医学部 獣医学科
職名 講師
researchmap研究者コード 5000068922
researchmap機関 岡山理科大学

題名

Kinase activity-tagged western blotting assay

単著・共著の別

共著

著者

Masumi Eto, Shuichi Katsuki, Yoshinori Tanaka, Kosuke Takeya

概要

Determining cellular activities of protein kinases is a fundamental step for characterizing pathophysiological cell signaling pathways. Here, we optimized a nonradioactive method that detects protein kinases in tissues or cells after separation by SDS-PAGE and transfer onto polyvinylidene fluoride membranes. The method, kinase activity-tagged western blotting (KAT-WB), consists of five steps: electrophoresis of cell extracts that contain protein kinases, electroblotting proteins onto polyvinylidene fluoride membrane, denaturation-renaturation, phosphorylation, with or without an added substrate protein and immunodetection using anti-phospho-specific antibodies. KAT-WB detected autophosphorylation of one Tyr-kinase and site-specific phosphorylation of added substrate by multiple kinases. KAT-WB assay enables us to interrogate multiple kinase signaling pathways without using radioactive ATP.    

発表雑誌等の名称

Biotechniques

出版者

68

4

開始ページ

211

終了ページ

213

発行又は発表の年月

2020/01

査読の有無

有り

招待の有無

無し

記述言語

英語

掲載種別

研究論文(学術雑誌)

ISSN

ID:DOI

10.2144/btn-2019-0136

ID:NAID(CiNiiのID)

ID:PMID

31939317

URL

JGlobalID

arXiv ID

ORCIDのPut Code

DBLP ID