JNK1/2/3 (Phospho-Thr183)Colorimetric Cell-Based ELISA Kit

The Assay Genie cell-based ELISA kit is designed for the sensitive and accurate detection of JNK1/2/3 phosphorylated at threonine 183 in cell lysates. The kit uses an indirect ELISA format in which the phospho-epitope is captured by JNK1/2/3 (Phospho-Thr183)-specific primary antibodies, and HRP-conjugated secondary antibodies bind the Fc region of the primary, catalysing a colorimetric reaction measured at 450 nm upon substrate addition. It is described by the manufacturer as a convenient, lysate-free, high-throughput, and sensitive assay that measures relative amounts of phosphorylated JNK1/2/3 in cultured cells, and can be used to screen the effects of treatments, inhibitors (including siRNA or chemicals), or activators on JNK1/2/3 phosphorylation.

Phosphorylation of JNK1/2/3 at threonine 183 is described as a key event in activation of the JNK signalling pathway, which regulates cellular processes including proliferation, apoptosis, and inflammation, and whose dysregulation has been implicated in cancer, inflammatory disorders, and neurodegenerative disease. The kit is reactive with human, mouse, and rat samples, is intended for adherent cells, and has a manufacturer-stated dynamic range of "> 5000 Cells". The target gene symbol is MAPK8/9/10 and the sub-type is phospho. 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 is provided so that, after the colorimetric readout, absorbance values can be normalised to cell amounts to adjust for plating differences.