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Real-Time Monitoring of Protein Degradation Using Bioluminescent Tagging and the GloMax® Discover

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Abstract

We show kinetic dose-response degradation curves for the bromodomain protein, BRD4, endogenously tagged using CRISPR/Cas9 with the peptide HiBiT in HEK293 cells stably expressing the LgBiT fragment. We used the GloMax® Discover microplate luminometer to continuously monitor BRD4 levels over 24 hours after treatment with the BRD4 PROTAC compound MZ1. Rapid degradation of endogenously tagged HiBiT-BRD4 was observed in a dose-dependent fashion with MZ1 PROTAC. At the highest concentration, almost complete degradation of BRD4 was obtained and sustained over the 24-hour period.

Application Note #AN342

Kristin Riching, Ph.D. Publication Date: 4/19

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