SHP-2 Colorimetric Cell-Based ELISA Kit

This Assay Genie cell-based ELISA kit offers a lysate-free, high-throughput workflow for profiling SHP-2 protein expression in cultured cells and for screening the effects of treatments, inhibitors (such as siRNA or chemicals) and activators. Detection uses a qualitative indirect ELISA format, in which SHP-2-specific primary antibodies are captured on the plate and HRP-conjugated secondary antibodies catalyse a colorimetric reaction read at 450 nm.

Reactivity has been confirmed for human, mouse and rat samples, with a stated dynamic range of > 5000 Cells. A monoclonal anti-GAPDH antibody is included as an internal positive control to normalise target absorbance, and a Crystal Violet whole-cell staining step is provided to correct for cell density after the colorimetric read. The standard protocol involves seeding 200 µL of 20,000 adherent cells per well, fixing with 4% formaldehyde, then proceeding through quenching, blocking, overnight primary antibody incubation at 4 °C, HRP-conjugated secondary incubation and substrate development before reading the plate.

SHP-2 is encoded by the PTPN11 gene (Gene ID: 5781; UniProt ID: Q06124), a ubiquitously expressed SH2-containing tyrosine-specific protein phosphatase that participates in signaling downstream of growth factor, cytokine, hormone, antigen and extracellular matrix receptors and contributes to the regulation of cell growth, differentiation, migration and death. The kit is stored at 4 °C with a stated stability of 6 months.