EFNA3 Colorimetric Cell-Based ELISA

From Assay Genie, this Cell-Based ELISA is designed for the precise measurement of EFNA3 (ephrin-A3) levels in cell lysates and tissue homogenates. The kit uses a colorimetric detection method at 450 nm and is reactive with human, mouse, and rat samples. It is described as a lysate-free, high-throughput, and sensitive assay suitable for measuring relative amounts of EFNA3 in cultured cells and for screening the effects of treatments, inhibitors, or activators on EFNA3 expression.

The assay employs an indirect ELISA format in which EFNA3 is captured by EFNA3-specific primary antibodies, while HRP-conjugated secondary antibodies bind the Fc region of the primary antibody. A monoclonal antibody specific for human GAPDH is included to serve as an internal positive control for normalising target absorbance values. Following the colorimetric measurement of HRP activity, the Crystal Violet whole-cell staining method may be used to determine cell density, allowing absorbance values to be normalised to cell amounts and plating differences to be adjusted.

EFNA3 is a cell surface GPI-bound ligand for Eph receptors, a family of receptor tyrosine kinases crucial for migration, repulsion, and adhesion during neuronal, vascular, and epithelial development, with roles in axon guidance, synaptogenesis, and angiogenesis. Dysregulation of EFNA3 has been implicated in cancer, neurological disorders, and cardiovascular conditions. The kit is supplied in a 96-Well Microplate format with a dynamic range of > 5000 cells, and should be stored at 4°C for 6 months.