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EndoBind detects endogenous protein-protein interactions in real time.

Anke BillSheryll EspinolaDaniel GuthyJacob R HalingMylene LanterMin LuAnthony MarelliAngelica MendiolaLoren J MiragliaBrandon L TaylorLeonardo VargasAnthony P OrthFrederick J King
Published in: Communications biology (2021)
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.
Keyphrases
  • high throughput
  • oxidative stress
  • single cell
  • real time pcr
  • amino acid
  • quantum dots