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Direct Measurement of KDM1A Target Engagement Using Chemoprobe-based Immunoassays
JoVE Journal
Biochemistry
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JoVE Journal Biochemistry
Direct Measurement of KDM1A Target Engagement Using Chemoprobe-based Immunoassays
DOI:

11:17 min

June 13, 2019

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Chapters

  • 00:04Title
  • 01:05Solution Preparation
  • 02:11Native Protein Extraction
  • 03:51Luminescent ELISAs for Total and Free KDM1A Determination
  • 08:42Results: Direct Measurement of KDM1A Target Engagement
  • 10:32Conclusion

Summary

Automatic Translation

Here, we present a protocol to measure KDM1A target engagement in a human or animal cell, tissue or blood samples treated with KDM1A inhibitors. The protocol employs chemoprobe tagging of the free KDM1A enzyme and direct quantification of the target occupation using chemoprobe-based immunoassays and can be used in preclinical and clinical studies.

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