SHP-2 (Phospho-Tyr542)Fluorometric Cell-Based ELISA Kit

Manufactured by Assay Genie, this fluorometric phospho-cell-based ELISA is designed to monitor phosphorylation of SHP-2 at tyrosine 542 in cultured cells. The assay uses a lysate-free, indirect ELISA format with fluorometric detection: phospho-SHP-2 (Tyr542) is captured by a target-specific primary antibody, and Dye 1- and Dye 2-conjugated secondary antibodies bound to the Fc region of the primary antibody emit light at defined wavelengths, allowing qualitative measurement of relative phosphorylated SHP-2 levels. The kit can be used to screen the effects of treatments, inhibitors (such as siRNA or chemicals) or activators on SHP-2 phosphorylation.

Two normalization approaches are built in: a monoclonal antibody against human GAPDH serves as an internal positive control for normalizing target RFU values, and a second antibody directed against the non-phosphorylated counterpart of SHP-2 is included so that RFU values for non-phosphorylated SHP-2 can normalize the readings for phosphorylated SHP-2. The dynamic range is reported as > 5000 cells, and the kit is reactive with Human, Mouse and Rat samples. Recommended storage is 4°C for 6 months. Fluorescence is read on a two-channel plate reader at Ex/Em 651/667 nm (Dye 1) and 495/521 nm (Dye 2).

The target corresponds to PTPN11 (Gene ID 5781; UniProt Q06124), a ubiquitously expressed SH2-containing tyrosine phosphatase implicated in signaling downstream of growth factor, cytokine and hormone receptors and in pathways controlling cell growth, differentiation, migration and death. Mutations in PTPN11 are associated with Noonan Syndrome 1, juvenile myelomonocytic leukemia, Leopard Syndrome 1 and metachondromatosis.