PKC alpha (Phospho-Thr638)Colorimetric Cell-Based ELISA Kit

From Assay Genie, this Cell-Based ELISA Kit is designed for the detection of phosphorylated PKC alpha (Thr638) levels in cultured cells. PKC alpha is a key signalling enzyme that regulates processes including cell growth, differentiation and apoptosis, and phosphorylation at the Thr638 site modulates its activity. Dysregulation of PKC alpha has been implicated in cancer, diabetes and neurological disorders, making its phosphorylation status a valuable target for therapeutic research.

The kit provides a convenient, lysate-free, high-throughput and sensitive assay that uses an indirect ELISA format: PKC alpha (Phospho-Thr638)-specific primary antibodies capture the target, while HRP-conjugated secondary antibodies catalyse a colorimetric reaction at 450 nm upon substrate addition. It can be used to measure the relative amounts of phosphorylated PKC alpha in cultured cells and to screen the effects of various treatments, inhibitors (such as siRNA or chemicals) or activators on PKC alpha phosphorylation. Reactivity is reported for human, mouse and rat samples, and the dynamic range is greater than 5000 cells. The intended sample type is adherent cells.

Because the assay is qualitative, two normalisation methods are provided. A monoclonal antibody specific for human GAPDH is included to serve as an internal positive control, and a Crystal Violet whole-cell staining step can be used to determine cell density after the colorimetric readout, allowing absorbance values to be normalised to cell amounts.