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Titolo:
Expression and purification of active recombinant ATM protein from transiently transfected mammalian cells
Autore:
Rhodes, N; Gilmer, TM; Lansing, TJ;
Indirizzi:
GlaxoSmithKline Res & Dev, Oncol Biol Dept, Res Triangle Pk, NC 27709 USA GlaxoSmithKline Res & Dev Res Triangle Pk NC USA 27709 e Pk, NC 27709 USA
Titolo Testata:
PROTEIN EXPRESSION AND PURIFICATION
fascicolo: 3, volume: 22, anno: 2001,
pagine: 462 - 466
SICI:
1046-5928(200108)22:3<462:EAPOAR>2.0.ZU;2-4
Fonte:
ISI
Lingua:
ENG
Soggetto:
ATAXIA-TELANGIECTASIA GENE; DNA-DAMAGE RESPONSE; DEPENDENT PHOSPHORYLATION; IONIZING-RADIATION; CHECKPOINT PATHWAY; KINASE-ACTIVITY; P53; ACTIVATION; WORTMANNIN; BRCA1;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
34
Recensione:
Indirizzi per estratti:
Indirizzo: Lansing, TJ GlaxoSmithKline Res & Dev, Oncol Biol Dept, 5 Moore Dr, Res Triangle Pk, NC 27709 USA GlaxoSmithKline Res & Dev 5 Moore Dr Res Triangle Pk NC USA 27709
Citazione:
N. Rhodes et al., "Expression and purification of active recombinant ATM protein from transiently transfected mammalian cells", PROT EX PUR, 22(3), 2001, pp. 462-466

Abstract

The gene mutated in the human disease ataxia telangiectasia (AT), termed ATM, encodes a large protein kinase involved in DNA repair and cell cycle control. Biochemical characterization of ATM function has been somewhat difficult because of its large size (similar to 370 kDa) and relatively low level of expression in several systems. The majority of studies have used immunoprecipitated ATM or purified ATM obtained through relatively complex procedures. Here, we describe an efficient method for the expression and purification of FLAG-epitope-tagged recombinant human ATM protein (F-ATM). This method utilizes the expression of F-ATM in transiently transfected 293T cellsfollowed by anti-FLAG-agarose affinity chromatography. The transfection procedure has been optimized for large (225-cm(2)) culture flasks and F-ATM can be purified to near homogeneity as judged by SDS-PAGE. This procedure yields approximately 1 mug of catalytically active F-ATM protein/225-cm(2) flask that can be used for biochemical studies. (C) 2001 Academic Press.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 02/06/20 alle ore 23:49:33