Shc (Phospho-Tyr349)Colorimetric Cell-Based ELISA Kit

A cell-based ELISA from Assay Genie designed for the accurate measurement of SHC phosphorylation at Tyrosine 349. The kit is a lysate-free, high-throughput and sensitive assay that can detect Shc protein phosphorylation and expression profile in cells, allowing researchers to measure the relative amounts of phosphorylated Shc in cultured cells and to screen for the effects of various treatments, inhibitors (such as siRNA or chemicals), or activators on Shc phosphorylation. Qualitative determination of Shc (Phospho-Tyr349) concentration is achieved by an indirect ELISA format in which the target is captured by Shc (Phospho-Tyr349)-specific primary antibodies while HRP-conjugated secondary antibodies bind the Fc region of the primary antibody; the conjugated HRP then catalyses a colorimetric reaction upon substrate addition. The assay is reactive with Human, Mouse and Rat samples, with a dynamic range of > 5000 Cells, detection by colorimetric measurement at 450 nm, and is suitable for adherent cells.

Two normalisation methods are provided: a monoclonal antibody specific for human GAPDH is included as an internal positive control for normalising target absorbance values, and a Crystal Violet whole-cell staining method may be used to determine cell density so that absorbance values can be normalised to cell amounts, adjusting for plating differences. SHC is a critical adaptor protein (gene symbol SHC1) containing an SH2 domain and a PID domain that couples activated growth factor receptors to downstream signalling pathways, with phosphorylation at Tyrosine 349 regulating its activity and function. Studying this phosphorylation event can offer insights into signalling pathways implicated in cancer, diabetes and other diseases, supporting research into cell signalling and the development of targeted therapies.