14-3-3 zeta Colorimetric Cell-Based ELISA Kit

Assay Genie offers a cell-based ELISA kit for the detection and quantification of 14-3-3 zeta protein levels in cell lysates and tissue lysates. The colorimetric detection method, measured at 450 nm, provides a lysate-free, high-throughput and sensitive assay for measuring relative amounts of 14-3-3 zeta in cultured cells, as well as for screening the effects of various treatments, inhibitors (such as siRNA or chemicals), or activators on the protein.

Detection is achieved through an indirect ELISA format in which 14-3-3 zeta is captured by specific primary antibodies, and HRP-conjugated secondary antibodies bind the Fc region of the primary antibody to catalyse a colorimetric reaction upon substrate addition. A monoclonal anti-GAPDH antibody is included as an internal positive control for normalising target absorbance values, while a Crystal Violet whole-cell staining method enables determination of cell density, allowing absorbance values to be normalised to cell amounts. The kit is reactive with human, mouse and rat samples and is intended for use with adherent cells, with a dynamic range of > 5000 cells and a 96-well microplate format. Storage is at 4°C with 6 months stability.

14-3-3 zeta (gene symbol YWHAZ) is an adapter protein implicated in cell cycle regulation, signal transduction and apoptosis. Dysregulation of the protein has been associated with cancer, neurodegenerative disorders and cardiovascular diseases, making it a relevant biomarker for research into cellular function and disease pathology.