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生物化学
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Identification of Novel CK2 Kinase Substrates Using a Versatile Biochemical Approach
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生物化学
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JoVE 杂志 生物化学
Identification of Novel CK2 Kinase Substrates Using a Versatile Biochemical Approach

Identification of Novel CK2 Kinase Substrates Using a Versatile Biochemical Approach

DOI:
10.3791/59037-v

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11:11 min

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February 21, 2019

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, ,
1Department of Biochemistry and Molecular Biology,Drexel University College of Medicine

Chapters

  • 00:04Title
  • 00:35Preparation of the Tissue Samples
  • 01:20Kinase Assay: Thiophosphorylation and Alkylation
  • 02:34Preparation of the Desalting Columns
  • 03:40Removal of PNBM
  • 04:42Immunoprecipitation: Part I
  • 05:32Protein A/G Agarose Bead Preparation
  • 06:39Immunoprecipitation: Part II
  • 08:19Analysis/Validation of Results
  • 09:06Results: Analysis and Identification of Novel CK2 Kinase Substrates
  • 10:48Conclusion

Summary

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The objective of this protocol is to label, enrich, and identify substrates of protein kinase CK2 from a complex biological sample such as a cell lysate or tissue homogenate. This method leverages unique aspects of CK2 biology for this purpose.

Tags

CK2Kinase SubstratesBiochemical ApproachEndogenous SubstratesCell LysateGTPgammaSPNBMSephadex G-25Affinity Purification

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