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Three optimized assays for the evaluation of compounds that can rescue p53 mutants

STAR Protoc. 2021-07; 
Jiale Wu, Huaxin Song, Zhengyuan Wang, Min Lu
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Custom DNA/RNA Oligos … ExpressPlus PAGE Gel, 10×8, 4–20%, GenScript, Cat# M42015C. White 96-well plates, SPL, Cat# 31396 … Alternatives: The DSF protocol describes the procedure using a LightCycler® 480 RT-PCR Detection system and corresponding reagents, in 10 μL reaction volumes … Get A Quote

摘要

Identifying drugs targeting p53 remains a major focus of precision oncology, with over twenty compounds that can rescue p53 mutants reported. Here, we suggest three easily accessible assays to determine the thermostability, protein folding, and transcriptional activity of p53 mutants-the go-to criteria for evaluating a rescue compound that acts by increasing p53 thermostability. Because of the diversity of p53 mutants, a compound that meets the criteria of one assay does not necessarily meet the criteria of the other assays. For complete details on the use and execution of this protocol, please refer to Chen et al. (2021).

关键词

Antibody, Cancer, Cell-based Assays, Protein Biochemistry