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High-Throughput Cellular Profiling of Targeted Protein Degradation Compounds Using HiBiT CRISPR Cell Lines
JoVE Journal
Ricerca sul cancro
This content is Free Access.
JoVE Journal Ricerca sul cancro
High-Throughput Cellular Profiling of Targeted Protein Degradation Compounds Using HiBiT CRISPR Cell Lines
DOI:

05:33 min

November 09, 2020

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Capitoli

  • 00:04Introduction
  • 00:50Endpoint Degradation Studies with HiBiT CRISPR Target Proteins in Lytic Format with Optional Cell Viability Fluorescence Analysis
  • 03:07Results: Degradation Compound Analysis in a HiBiT CRISPR Cell Line
  • 04:51Conclusion

Summary

Traduzione automatica

This protocol describes the quantitative luminescent detection of protein degradation kinetics in living cells that have been engineered using CRISPR/Cas9 to express antibody free endogenous protein detection tag fused to a target protein. Detailed instructions for calculating and obtaining quantitative degradation parameters, rate, Dmax, DC50, and Dmax50 are included.

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