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EndoBind detects endogenous protein-protein interactions in real time

Commun Biol. 2021-09;

Anke Bill, Sheryll Espinola, Daniel Guthy, Jacob R Haling, Mylene Lanter, Min Lu, Anthony Marelli, Angelica Mendiola, Loren Miraglia, Brandon L Taylor, Leonardo Vargas, Anthony P Orth, Frederick J King

Products/Services Used	Details	Operation
Gene Synthesis	… All gene synthesis and cloning were performed by Genscript. Full sequences of the inserts can be found in the source data. The sgRNAs targeting the integration sites were cloned into a modified U6-based puromycin-resistant pLKO vector, including an expression cassette for …	Get A Quote

摘要

We present two high-throughput compatible methods to detect the interaction of ectopically expressed (RT-Bind) or endogenously tagged (EndoBind) proteins of interest. Both approaches provide temporal evaluation of dimer formation over an extended duration. Using examples of the Nrf2-KEAP1 and the CRAF-KRAS-G12V interaction, we demonstrate that our method allows for the detection of signal for more than 2 days after substrate addition, allowing for continuous monitoring of endogenous protein-protein interactions in real time.