PLCG1 Colorimetric Cell-Based ELISA Kit

From Assay Genie, this cell-based ELISA kit is designed for the qualitative detection of PLCG1 protein expression in cultured cells. It is reactive with human, mouse, and rat samples, uses a colorimetric readout measured at 450 nm, and is supplied in a microplate format with a stated dynamic range of > 5000 cells. The indicated sample type is adherent cells.

The assay follows an indirect ELISA format: PLCG1 is captured by target-specific primary antibodies and detected with HRP-conjugated secondary antibodies, which catalyze a colorimetric reaction upon substrate addition. Because the readout is qualitative, two normalization methods are included: a monoclonal anti-GAPDH antibody serves as an internal positive control, and a Crystal Violet whole-cell staining step is used to determine cell density so absorbance values can be adjusted for plating differences. The procedure involves seeding cells, fixing, quenching, blocking, overnight primary antibody incubation at 4 °C (shortened to two hours at room temperature when target expression is known to be high), secondary antibody incubation, and color development.

PLCG1 is described as a calcium-dependent phosphatidylinositol-specific phospholipase C that generates the second messengers diacylglycerol and inositol 1,4,5-trisphosphate and is phosphorylated and activated by tyrosine kinases in response to signaling through various growth factor and immune system receptors. The kit is stored at 4 °C and is stated to be stable for 6 months under those conditions.