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Identification of Cyclin-dependent Kinase 1 Specific Phosphorylation Sites by an In Vitro Kinase Assay
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Journal JoVE Biochimie
Identification of Cyclin-dependent Kinase 1 Specific Phosphorylation Sites by an In Vitro Kinase Assay

Identification of Cyclin-dependent Kinase 1 Specific Phosphorylation Sites by an In Vitro Kinase Assay

DOI:
10.3791/57674-v

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12:26 min

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May 03, 2018

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, , ,
1Department of Chemistry,State University of New York at Binghamton

Chapitres

  • 00:04Titre
  • 01:25Prediction of Cdk1-specific Phosphorylation Sites and Expression of CENP-F in Escherichia coli
  • 04:11Recombitant Protein Purification by Glutathione-affinity Chromatography and Gel Filtration Followed by In Vitro Kinase Assay with Cdk1/Cyclin B
  • 09:50Results: Use of In Vitro Kinase Assay to Study Cdk1-specific Phosphorylation Sites in CENP-F
  • 11:13Conclusion

Summary

Traduction automatique

Traduction automatique

Cyclin-dependent kinase 1 (Cdk1) is activated in the G2 phase of the cell cycle and regulates many cellular pathways. Here, we present a protocol for an in vitro kinase assay with Cdk1, which allows the identification of Cdk1-specific phosphorylation sites for establishing cellular targets of this important kinase.

Tags

CDK1PhosphorylationKinase AssayCell CycleCENP-FProtein ExpressionE. ColiLB MediumAntibioticsProtein Purification

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