PKC alpha (Phospho-Tyr657)Colorimetric Cell-Based ELISA Kit

A cell-based ELISA from Assay Genie for the qualitative detection of phosphorylated PKC alpha at tyrosine 657 directly in cultured cells. The assay is lysate-free and uses an indirect ELISA format, in which phospho-specific primary antibodies capture the target and HRP-conjugated secondary antibodies generate a colorimetric signal at 450 nm upon substrate addition.

Designed for use with adherent cells and reporting a dynamic range of > 5000 Cells, the kit is reactive with human, mouse, and rat samples. It can be used to measure the relative phosphorylation status of PKC alpha in cultured cells and to screen the effects of treatments, inhibitors (including siRNA or chemicals), or activators on PKC alpha phosphorylation. A monoclonal anti-GAPDH antibody is included as an internal positive control to normalize target absorbance values, and a Crystal Violet whole-cell staining method is provided so that absorbance readings can be adjusted to cell density and plating differences.

The target is encoded by PRKCA (Gene ID: 5578; UniProt ID: P17252), a classical calcium-dependent AGC kinase of the PKC family. PKC-Alpha is a key regulator of cell growth, differentiation, and apoptosis, and phosphorylation at tyrosine 657 plays a critical role in its activation and downstream function. The kit is suited to applications including the study of signal transduction pathways, kinase signaling cascades, drug discovery, cancer research, and molecular biology.