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Titolo:
HARVEST PROTOCOL TO REDUCE VARIABILITY OF SOLUBLE ENZYME YIELD FROM CULTURED-CELLS
Autore:
BUSKIN JN; GREGORY DL; LAFRAMBOISE WA; HAUSCHKA SD;
Indirizzi:
UNIV WASHINGTON,DEPT BIOCHEM,BOX 357350 SEATTLE WA 98195
Titolo Testata:
BioTechniques
fascicolo: 1, volume: 20, anno: 1996,
pagine: 92 -
SICI:
0736-6205(1996)20:1<92:HPTRVO>2.0.ZU;2-0
Fonte:
ISI
Lingua:
ENG
Soggetto:
CREATINE-KINASE ENHANCER; MUSCLE; SKELETAL; BINDING; GENE;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
10
Recensione:
Indirizzi per estratti:
Citazione:
J.N. Buskin et al., "HARVEST PROTOCOL TO REDUCE VARIABILITY OF SOLUBLE ENZYME YIELD FROM CULTURED-CELLS", BioTechniques, 20(1), 1996, pp. 92

Abstract

Many aspects of physiology and gene regulation can be studied by examining the levels of enzymes harvested from cultured cells. We found that the yield from cultured cells of two different cytosolic enzymes, creatine kinase and the common reporter gene product chloramphenicol acetyltransferase (CAT), could be highly variable despite superficially identical harvest procedures. Analysis of multiple harvest and assay parameters disclosed that fluctuations in enzyme yield were correlated with the rime cells that were allowed to remain in an EDTA-containing buffered saline solution prior to scraping from the dishes with a rubber policeman. The highest and most consistent yields were obtained when the cells were allowed to remain in the solution for 6-10 min beforescraping; this protocol has cut variability approximately by a factorof three.

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Documento generato il 26/09/20 alle ore 13:12:41