PKC theta (Phospho-Thr538)Colorimetric Cell-Based ELISA Kit

From Assay Genie, this colorimetric cell-based ELISA kit is designed for the detection of phosphorylated PKC theta at Thr538 in cell lysates and culture supernatants. The assay targets the Thr538 phosphorylation event, a key regulatory modification of PKC theta that influences T cell activation and differentiation, making the kit relevant to research into immune cell function, autoimmune disease, and related therapeutic targets.

The kit uses an indirect ELISA format and is lysate-free, allowing measurement of relative phosphorylated PKC theta levels directly in cultured cells. It is compatible with adherent cell samples and is reactive with human, mouse, and rat samples. Detection is carried out by reading absorbance at 450 nm, with a dynamic range supporting assays using more than 5000 cells. A monoclonal anti-GAPDH antibody is included as an internal positive control for normalization, and a Crystal Violet whole-cell staining step can be used to determine cell density and adjust for plating differences. The kit is suitable for screening the effects of treatments, inhibitors (such as siRNA or chemical compounds), or activators on PKC theta phosphorylation, with HRP-conjugated secondary antibodies catalysing the colorimetric reaction upon substrate addition.

PKC theta is encoded by the PRKCQ gene (Gene ID 5588; UniProt Q04759; OMIM 600448) and belongs to the AGC kinase family. It is a calcium-independent, phospholipid-dependent serine/threonine kinase involved in T cell receptor signalling and NF-kappaB activation, and is expressed in skeletal muscle and haematopoietic cells.