PYK2 (Phospho-Tyr580)Colorimetric Cell-Based ELISA Kit

From Assay Genie, this cell-based ELISA kit is designed for the measurement of phosphorylated PYK2 (phospho-Tyr580) levels in cultured cells. The assay uses a colorimetric detection method read at 450 nm and is reactive with human, mouse, and rat samples. It is intended for use with adherent cells in a lysate-free workflow, and a dynamic range of > 5000 cells is reported.

The kit employs an indirect ELISA format in which phospho-PYK2 is captured by target-specific primary antibodies, with HRP-conjugated secondary antibodies binding the Fc region of the primaries to catalyse the colorimetric reaction upon substrate addition. A monoclonal antibody specific for human GAPDH is included as an internal positive control in normalising target absorbance values, and a Crystal Violet whole-cell staining step can be used to determine cell density so that absorbance values can be adjusted to cell amounts, correcting for plating differences. The qualitative approach allows measurement of relative phosphorylated PYK2 in cultured cells and screening of the effects that various treatments, inhibitors (such as siRNA or chemicals), or activators have on PYK2 phosphorylation.

PYK2, also known as protein tyrosine kinase 2 beta, is a non-receptor tyrosine kinase of the FAK family. It is encoded by the PTK2B gene (Gene ID 2185, UniProt ID Q14289, OMIM 601212, Unigene Hs.491322), and two alternatively spliced isoforms have been described. The sub-type of the kit is Phospho, with PYK2 (Phospho-Tyr580) as the target.