Scientific BackgroundThought to form a receptor-activated non-selective calcium permeant cation channel (PubMed:19936226, PubMed:23291369). Probably is operated by a phosphatidylinositol second messenger system activated by receptor tyrosine kinases or G-protein coupled receptors. Activated by diacylglycerol (DAG) in a membrane-delimited fashion, independently of protein kinase C (PubMed:26892346). Seems not to be activated by intracellular calcium store depletion (uniprot).
A. Western Blot: (1 µg/mL) staining of A549 cell lysate (35 ug protein in RIPA buffer). Detected by chemiluminescence. B. Flow cytometry analysis: of paraformaldehyde fixed A549 cells (blue line), permeabilized with 0.5% Triton. Primary incubation 1hr (10 µg/mL) followed by Alexa Fluor 488 secondary antibody (1 µg/mL). IgG control: Unimmunized goat IgG (black line) followed by Alexa Fluor 488 secondary antibody.