JAK2 (Phospho-Tyr221)Colorimetric Cell-Based ELISA Kit

From Assay Genie, this Cell-Based ELISA Kit is designed to detect phosphorylation of JAK2 at tyrosine 221 in cultured cells. It is a convenient, lysate-free, high throughput and sensitive assay that can be used to measure the relative amounts of phosphorylated JAK2 in cultured cells and to screen for the effects that various treatments, inhibitors (such as siRNA or chemicals), or activators have on JAK2 phosphorylation. The kit shows reactivity with Human, Mouse, and Rat samples and operates with a dynamic range of > 5000 Cells.

Detection is achieved through a colorimetric method at 450 nm, using an indirect ELISA format in which JAK2 (Phospho-Tyr221) is captured by JAK2 (Phospho-Tyr221)-specific primary antibodies and HRP-conjugated secondary antibodies catalyse a colour reaction upon substrate addition. A monoclonal anti-GAPDH antibody is included as an internal positive control for normalising target absorbance values, and Crystal Violet whole-cell staining can be used to determine cell density, allowing absorbance values to be normalised to cell amounts and adjusted for plating differences.

The target, JAK2, is a non-receptor tyrosine kinase involved in a specific subset of cytokine receptor signaling pathways, including IL-3, IL-5 and GM-CSF, and plays a key role in the JAK-STAT signaling pathway. Phosphorylation of JAK2 at tyrosine 221 is known to regulate its activity, making the kit relevant for studies of signal transduction, immune response, cell proliferation and related physiological and pathological processes.