SHP-2 (Phospho-Tyr580)Colorimetric Cell-Based ELISA Kit

Designed for the detection of SHP-2 phosphorylation at tyrosine 580 in cell samples, this Colorimetric Cell-Based ELISA Kit from Assay Genie provides a convenient, lysate-free, high-throughput and sensitive assay that can detect SHP-2 protein phosphorylation and expression profile in cells, and is intended for use with adherent cells.

Qualitative determination is achieved by an indirect ELISA format in which the target is captured by SHP-2 (Phospho-Tyr580)-specific primary antibodies, with HRP-conjugated secondary antibodies binding the Fc region of the primary antibody and catalysing a colorimetric reaction upon substrate addition, measured at 450 nm. The assay is reactive with Human, Mouse and Rat samples and has a dynamic range of > 5000 Cells. A monoclonal antibody specific for human GAPDH is included to serve as an internal positive control for normalising the target absorbance values, and a Crystal Violet whole-cell staining method may be used to determine cell density, after which absorbance values can be normalised to cell amounts to adjust for plating differences.

SHP-2 is a ubiquitously expressed, SH2-containing tyrosine-specific protein phosphatase that participates in signalling downstream of receptors for growth factors, cytokines, hormones, antigens and extracellular matrices, regulating cell growth, differentiation, migration and survival. Phosphorylation at tyrosine 580 modulates its activity and function, impacting pathways implicated in cancer, metabolic disorders and immune responses. The kit can be used to measure the relative amounts of phosphorylated SHP-2 in cultured cells and to screen for the effects that various treatments, inhibitors (such as siRNA or chemicals) or activators have on SHP-2 phosphorylation.