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A tandem chromatographic column method for assaying cAMP-dependent protein kinase and protein kinase C with synthetic peptide substrates

✍ Scribed by John J. Egan; Min-Kun Chang; Constantine Londos


Publisher
Elsevier Science
Year
1988
Tongue
English
Weight
878 KB
Volume
175
Category
Article
ISSN
0003-2697

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✦ Synopsis


A method was devised for assaying protein kinases that phosphorylate either Kemptide, such as cAMP-dependent protein kinase, or a glycogen synthase peptide, which is an excellent substrate for protein kinase C. Upon sequential processing of reaction mixtures through tandem columns of cation and anion exchange resins, radioactivity in background samples is nearly nil and the yield of phosphorylated peptides is high. This method reduces labor, radioactivity, enzyme requirements, and costs of assaying protein kinases.