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Co-immunoprecipitation Assay for Studying Functional Interactions Between Receptors and Enzymes
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Co-immunoprecipitation Assay for Studying Functional Interactions Between Receptors and Enzymes
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Co-immunoprecipitation Assay for Studying Functional Interactions Between Receptors and Enzymes

DOI:
10.3791/58433-v

•

09:40 min

•

September 28, 2018

•

, ,
1Pulmonary Department,The Chaim Sheba Medical Center, 2Columbia Center for Translational Immunology, Department of Medicine,Columbia University Medical Center

Chapters

  • 00:04Title
  • 00:37Transfection of Cells
  • 01:54Promoting Phosphorylation in Transfection Cells
  • 02:40Immunoprecipitation
  • 06:13Phosphatase Activity Assay
  • 07:07SHP2 Western Blot Analysis
  • 08:16Results: Analysis of Co-immunoprecipitation Assay
  • 09:18Conclusion

Summary

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Here, we present a protocol for co-immunoprecipitation and an on-bead enzymatic activity assay to simultaneously study the contribution of specific protein domains of plasma membrane receptors to both enzyme recruitment and enzyme activity.

Tags

Co-immunoprecipitationReceptor-enzyme InteractionCell SignalingProtein DomainsEnzymatic ActivationDMEM293-T CellsTransfectionGFP ExpressionPervanadatePhosphorylationPD-1Lysis BufferProtease InhibitorsSodium OrthovanadateImmunoprecipitation

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