Method of detection of protein kinase activity in polyacrylamide gel using two-dimensional protein separating system

Method for detection of protein kinase activity in polyacrylamide gel have been developed. After separation of proteins by isoelectric focusing in non-denaturing condition, gel was incubated in a reaction buffer containing [gamma-32P]ATP. 32P-labeled proteins were separated by subsequent SDS/PAGE el...

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Bibliographic Details
Published inUkrains'kyi biokhimichnyi zhurnal (1999 ) Vol. 74; no. 4; p. 129
Main Authors Kit, Iu Ia, Koval'ova, V A, Bobak, Ia P, Drobot, L B
Format Journal Article
LanguageUkrainian
Published Ukraine 01.07.2002
Subjects
Animals
Blood Proteins - analysis
Drosophila melanogaster - enzymology
Drosophila Proteins - analysis
Electrophoresis, Gel, Two-Dimensional - methods
Humans
Isoelectric Focusing - methods
Protein Kinases - analysis
Protein Kinases - blood
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Summary:Method for detection of protein kinase activity in polyacrylamide gel have been developed. After separation of proteins by isoelectric focusing in non-denaturing condition, gel was incubated in a reaction buffer containing [gamma-32P]ATP. 32P-labeled proteins were separated by subsequent SDS/PAGE electrophoresis in second dimension. The proposed method was used for detection of protein kinase activity in human blood serum and triton X-100 soluble proteins of heads of Drosophila melanogaster.