Kinase Purification Resources and General Immunoprecipitation Pipeline
1:49
Kinase Reactions and SDS-PAGE
2:45
Gel Staining and Drying
4:18
Autoradiogram and Scintillation Counts
5:57
Results: Differences in Activity Between ERK2 and ERK2 Mutants
7:02
Conclusion
Transcript
The overall goal of this assay is to measure the enzymatic activity of protein kinases. This method can help answer key questions in the kinase signaling field, such as how active a kinase or kinase mutant is, its specificity, and whether its acti
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Protein kinases are highly evolved signaling enzymes and scaffolds that are critical for inter- and intracellular signal transduction. We present a protocol for measuring kinase activity through the use of radiolabeled adenosine triphosphate ([γ-32P] ATP), a reliable method to aid in elucidation of cellular signaling regulation.