IRS-1 (Phospho-Ser312)Colorimetric Cell-Based ELISA Kit

From Assay Genie, this Colorimetric Cell-Based ELISA Kit is designed for the accurate detection of phosphorylated IRS-1 at serine 312 in cell lysates. The kit is a convenient, lysate-free, high throughput and sensitive assay that can detect IRS-1 protein phosphorylation and expression profile in cells, making it suitable for measuring the relative amounts of phosphorylated IRS-1 in cultured cells and for screening the effects of various treatments, inhibitors, or activators on IRS-1 phosphorylation.

Qualitative determination of IRS-1 (Phospho-Ser312) is achieved by an indirect ELISA format, in which IRS-1 (Phospho-Ser312)-specific primary antibodies are captured while HRP-conjugated secondary antibodies bind the Fc region of the primary antibody, catalysing a colorimetric reaction upon substrate addition. A monoclonal antibody specific for human GAPDH is included as an internal positive control for normalisation, and a Crystal Violet whole-cell staining method may be used to determine cell density, allowing absorbance values to be normalised to cell amounts. The kit is reactive with Human, Mouse, and Rat samples, has a dynamic range of > 5000 Cells, and uses a colorimetric detection method at 450 nm.

Phosphorylation of IRS-1 at serine 312 plays a critical role in insulin signalling pathways and metabolic regulation, and its dysregulation has been linked to insulin resistance, diabetes, and other metabolic disorders, making it a relevant biomarker for research in these areas. The assay is intended for use with adherent cells.