FAK Colorimetric Cell-Based ELISA Kit

A colorimetric Cell-Based ELISA Kit from Assay Genie designed for the qualitative detection of FAK (focal adhesion kinase 1; gene symbol PTK2) in cultured cells. The assay uses an indirect ELISA format in which FAK is captured by FAK-specific primary antibodies and detected via HRP-conjugated secondary antibodies; following substrate addition, HRP catalyses a colorimetric reaction measured at 450 nm. Reactivity is reported for Human, Mouse, and Rat samples.

The kit is lysate-free and intended for high-throughput measurement of relative FAK levels in adherent cells, as well as for screening the effects of treatments, inhibitors (such as siRNA or chemicals), or activators on FAK. A monoclonal antibody specific for human GAPDH is included to serve as an internal positive control for normalising target absorbance values, and a Crystal Violet whole-cell staining step can be used to determine cell density and adjust for plating differences. The dynamic range is > 5000 Cells. The plate format is a 96-well microplate, and storage/stability is 4°C for 6 months.

FAK is a cytoplasmic protein tyrosine kinase of the FAK family that localises to focal adhesions formed between cells growing on extracellular matrix. It is activated by tyrosine phosphorylation in response to integrin clustering, GPCR occupancy by ligands such as bombesin or lysophosphatidic acid, or LDL receptor occupancy, acting downstream of integrins and Src and upstream of Ras/MAPK. FAK participates in cell motility, proliferation, and contact-dependent survival signalling. Increased expression has been reported in breast cancer (associated with chromosome 8q amplification), as well as in ovarian, hepatocellular, thyroid, and acute myelogenous leukaemia contexts; four splice-variant isoforms have been described.