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Identifikation af roman CK2 Kinase substrater ved hjælp af en alsidig biokemiske tilgang
JoVE Journal
Biochemistry
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JoVE Journal Biochemistry
Identification of Novel CK2 Kinase Substrates Using a Versatile Biochemical Approach
DOI:

11:11 min

February 21, 2019

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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

Automatic Translation

Formålet med denne protokol er at mærke, berige og identificere substrater af protein kinase CK2 fra en kompleks biologisk prøve celle lysate f.eks væv homogenatet. Denne metode udnytter unikke aspekter af CK2 biologi til dette formål.

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