JAK3 (Phospho-Tyr785)Colorimetric Cell-Based ELISA Kit

Manufactured by Assay Genie, this Cell-Based ELISA Kit is designed for the accurate measurement of JAK3 phosphorylation at tyrosine 785 in cell lysates. The assay uses a colorimetric detection method at 450 nm in an indirect ELISA format, in which a phospho-specific primary antibody captures JAK3 (Phospho-Tyr785) and an HRP-conjugated secondary antibody binds the Fc region of the primary antibody. The HRP enzyme conjugated to the secondary antibody then catalyses a colorimetric reaction upon substrate addition, enabling qualitative determination of JAK3 (Phospho-Tyr785) levels.

The kit is described as convenient, lysate-free, high throughput, and sensitive, and can detect JAK3 protein phosphorylation and expression profiles in cells. It is suited to measuring the relative amounts of phosphorylated JAK3 in cultured cells, as well as to screening the effects of various treatments, inhibitors such as siRNA or chemicals, or activators on JAK3 phosphorylation. Reactivity is reported for Human, Mouse, and Rat samples, and the dynamic range is given as "> 5000 Cells". The product is intended for use with adherent cells.

For data normalization, a monoclonal antibody specific for human GAPDH is included as an internal positive control, and the Crystal Violet whole-cell staining method can be used to determine cell density, allowing absorbance values to be normalized to cell amounts and plating differences to be adjusted. JAK3 (Janus kinase 3) is a non-receptor tyrosine-kinase of the Jak family, predominantly expressed in immune cells, that mediates signal transduction via the common gamma-chain of cytokine receptors including IL-2, -4, -7, -9, and -15. Phosphorylation of JAK3 at tyrosine 785 is known to play a key role in its activation and downstream signaling cascades, and dysregulation of this phosphorylation has been implicated in autoimmune disorders and certain cancers.