PI3-kinase p85-alpha/gamma (Phospho-Tyr467/199)Colorimetric Cell-Based ELISA Kit

A cell-based ELISA kit from Assay Genie designed to detect phosphorylation of PI3-kinase p85-alpha/gamma at tyrosine residues 467/199 in cell lysates. The assay uses colorimetric detection at 450 nm and is intended for use with adherent cell samples, with reactivity demonstrated against human, mouse, and rat samples. It is suited to research on the PI3 Kinase signaling pathway, including studies relevant to cancer, diabetes, and inflammatory disease, and provides a dynamic range of > 5000 Cells.

The kit employs an indirect ELISA format in which PI3-kinase p85-alpha/gamma (Phospho-Tyr467/199)-specific primary antibodies capture the target, while HRP-conjugated secondary antibodies bind the Fc region of the primary antibody, allowing the HRP enzyme to catalyze a colorimetric reaction upon substrate addition. A monoclonal antibody specific for human GAPDH is included to serve as an internal positive control for normalizing target absorbance values, and Crystal Violet whole-cell staining can be used to determine cell density so that absorbance values can be adjusted for plating differences. The assay is lysate-free and high-throughput, enabling measurement of relative phosphorylation levels in cultured cells and screening of the effects of various treatments, inhibitors, or activators on PI3-kinase p85-alpha/gamma phosphorylation.