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Utilizing a Comprehensive Immunoprecipitation Enrichment System to Identify an Endogenous Post-translational Modification Profile for Target Proteins
JoVE Journal
Biochemistry
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JoVE Journal Biochemistry
Utilizing a Comprehensive Immunoprecipitation Enrichment System to Identify an Endogenous Post-translational Modification Profile for Target Proteins

Utilizing a Comprehensive Immunoprecipitation Enrichment System to Identify an Endogenous Post-translational Modification Profile for Target Proteins

DOI:
10.3791/56912-v

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08:12 min

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January 08, 2018

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, ,
1R&D Department,Cytoskeleton Inc.

Chapters

  • 00:05Title
  • 00:59Sample Preparation: Cell Culture
  • 02:56Immunoprecipitation (IP) Assay
  • 07:00Results: Sample Preparation and IP Enrichment for PTM Detection and Crosstalk Investigation
  • 07:43Conclusion

Summary

Automatic Translation

Automatic Translation

Investigating multiple, endogenous post-translational modifications for a target protein can be extremely challenging. Techniques described here utilize an optimized lysis buffer and filter system developed with specific PTM-targeting, affinity matrices to detect the acetylation, ubiquitination, SUMOylation 2/3, and tyrosine phosphorylation modifications for a target protein using a single, streamlined system.

Tags

Keywords: ImmunoprecipitationPTMPost-translational ModificationCell SignalingLysateBlaster FilterProtein QuantitationA431 CellsLysis BufferDNA Removal

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