PKC theta (Phospho-Ser695)Colorimetric Cell-Based ELISA Kit

Assay Genie's cell-based ELISA kit is designed to detect phosphorylated PKC theta protein at the Ser695 site in cultured cells. The kit uses a colorimetric detection method read at 450 nm and is configured for human samples, with a dynamic range of > 5000 cells. It operates as a lysate-free, high-throughput assay that qualitatively measures relative amounts of phosphorylated PKC theta in cells, offering high sensitivity and specificity for research use.

The workflow follows an indirect ELISA format: the target is captured by PKC theta (Phospho-Ser695)-specific primary antibodies, and HRP-conjugated secondary antibodies bind the Fc region of the primary antibody, catalysing a colorimetric reaction upon substrate addition. A monoclonal anti-GAPDH antibody is included to serve as an internal positive control for normalising target absorbance values, and a Crystal Violet whole-cell staining step may be used to determine cell density so that absorbance values can be adjusted for plating differences.

PKC theta is a calcium-independent, phospholipid-dependent member of the protein kinase C family that is expressed in skeletal muscle and haematopoietic cells, and is required for T cell receptor signalling and NF-kappaB activation in mature T lymphocytes. The kit is suited to research on immune and inflammatory responses, autoimmune disorders, cancer and infectious diseases, and can be used to screen the effects of treatments, inhibitors (such as siRNA or chemical compounds) and activators on PKC theta phosphorylation.