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Cancer Research

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JoVE Journal Cancer Research
High-Throughput Cellular Profiling of Targeted Protein Degradation Compounds Using HiBiT CRISPR Cell Lines
 

High-Throughput Cellular Profiling of Targeted Protein Degradation Compounds Using HiBiT CRISPR Cell Lines

Article DOI: 10.3791/61787-v • 05:33 min • November 9th, 2020
November 9th, 2020 •
1, 1, 1, 1
1Promega Corporation

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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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High-throughput Cellular Profiling Targeted Protein Degradation HiBiT CRISPR Cell Lines Degradation Dynamics Sensitive And Quantitative Monitoring Degradation Parameters HTS Profiling Degrader Compound Activity Early Therapeutic Discovery Phase Modulation Of Endogenous Targets Celia Bisbach Mammalian Adherent Cells Suspension Cells Cell Density Serially-diluted PROTAC Degrader Test Compound Plates DMSO Control Incubation Endpoint Luminescent Signal Lytic Detection Reagent
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