A BRET Ca²⁺ sensor enables high-throughput screening in the presence of background fluorescence.

ABSTRACT: The intrinsic fluorescence of samples confounds the use of fluorescence-based sensors. This is of particular concern in high-throughput screening (HTS) applications using large chemical libraries containing intrinsically fluorescent compounds. To overcome this problem, we developed a bioluminescence resonance energy transfer (BRET) Ca²⁺ sensor, CalfluxCTN. We demonstrated that it reliably reported changes in intracellular Ca²⁺ concentrations evoked by an agonist and an antagonist of the human muscarinic acetylcholine receptor M₁ (hM₁R) even in the presence of the fluorescent compound fluorescein, which interfered with a standard fluorescent HTS sensor (Fluo-8). In an HTS using a chemical library containing fluorescent compounds, CalfluxCTN accura

SUBMITTER: Cumberbatch D 

PROVIDER: S-EPMC9930640 | biostudies-literature | 2022 Aug

REPOSITORIES: biostudies-literature