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Analyzing the regulation and function of ATM

Citation

Lavin, MF and Scott, SP and Kozlov, S and Gueven, N, Analyzing the regulation and function of ATM, Methods in Molecular Biology, 281 pp. 163-178. ISSN 1064-3745 (2004) [Refereed Article]

Copyright Statement

Copyright 2004 Humanities Press Inc.

Official URL: http://link.springer.com/protocol/10.1385/1-59259-...

Abstract

We describe here the cloning of full-length ataxia-telangiectasia mutated (ATM) cDNA and characterization of its activity. Full-length ATM cDNA is cloned into an inducible EBV-based vector (pMEP4) and its expression analyzed in a stably transfected cell line. ATM protein induction is monitored by immunoblotting with antibodies against both ATM and a FLAG sequence tag in the recombinant protein. Extracts from irradiated cells are immunoprecipitated with anti-ATM antibodies, and protein kinase activity is measured using p53(1-44)-specific substrate or by immunoblotting extracts with an anti-phosphoserine 15 p53-specific antibody. Missense mutations affecting ATM kinase activity are detected using in vitro mutagenesis of ATM cDNA followed by the procedures outlined above.

Item Details

Item Type:Refereed Article
Keywords:ATM, kinase
Research Division:Biological Sciences
Research Group:Biochemistry and Cell Biology
Research Field:Enzymes
Objective Division:Expanding Knowledge
Objective Group:Expanding Knowledge
Objective Field:Expanding Knowledge in the Biological Sciences
Author:Gueven, N (Dr Nuri Guven)
ID Code:79320
Year Published:2004
Deposited By:Pharmacy
Deposited On:2012-08-31
Last Modified:2014-11-05
Downloads:0

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