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Assaying Protein Kinase Activity with Radiolabeled ATP
JoVE Journal
Bioquímica
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JoVE Journal Bioquímica
Assaying Protein Kinase Activity with Radiolabeled ATP
DOI:

08:05 min

May 26, 2017

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Capítulos

  • 00:05Título
  • 00:36Kinase Purification Resources and General Immunoprecipitation Pipeline
  • 01:49Kinase Reactions and SDS-PAGE
  • 02:45Gel Staining and Drying
  • 04:18Autoradiogram and Scintillation Counts
  • 05:57Results: Differences in Activity Between ERK2 and ERK2 Mutants
  • 07:02Conclusion

Summary

Tadução automática

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.

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