IL-7R/CD127 (Phospho-Tyr449)Colorimetric Cell-Based ELISA Kit

From Assay Genie, this Cell-Based ELISA Kit is designed to detect phosphorylation of IL-7R/CD127 at tyrosine 449 in cultured cells. It provides a lysate-free, high throughput and sensitive approach for the qualitative determination of IL-7R/CD127 (Phospho-Tyr449) in cell-based assays, and can be used for measuring the relative amounts of phosphorylated IL-7R/CD127 as well as screening for the effects that various treatments, inhibitors (such as siRNA or chemicals), or activators have on IL-7R/CD127 phosphorylation. The kit shows reactivity with human and mouse samples and is intended for use with adherent cells.

The assay uses an indirect ELISA format in which IL-7R/CD127 (Phospho-Tyr449) is captured by specific primary antibodies, with HRP-conjugated secondary antibodies binding the Fc region of the primary antibody. Colour is developed via a colorimetric reaction at 450 nm upon substrate addition. A monoclonal antibody specific for human GAPDH is included as an internal positive control in normalizing the target absorbance values. Following the colorimetric measurement of HRP activity via substrate addition, a Crystal Violet whole-cell staining method may be used to determine cell density, allowing absorbance values to be normalized to cell amounts to adjust for plating differences. The dynamic range is reported as greater than 5000 cells.

By measuring phosphorylation of IL-7R/CD127 at tyrosine 449, the kit supports investigation of intracellular signalling events that regulate immune cell function, activation, proliferation and differentiation.