Application Brief 1014: PLC gamma-1 NanoPro Assay

Phospholipase C (PLC) catalyzes the hydrolysis of phosphatidylinositol 4, 5-bisphosphate (PIP2) to produce the metabolite second messenger molecules inositol 1,4,5-trisphosphate (IP3) and diacylglycerol (DAG). Increase of IP3 results in elevated intracellular free Ca²⁺. PLC's are activated through G-protein coupled receptor stimulation as well as tyrosine receptor kinase activation and therefore bridge both important signaling pathways. The family of PLC's consists of 12 isoforms with different roles in signaling. For example, PLCγ1 forms a complex with activated EGF receptors, which leads to the phosphorylation of PLCγ at Tyr771, 783 and 1245. Here we detect the phosphorylation of PLCγ1 in HEK293 cells in response to EGF treatment.