GADD153 Colorimetric Cell-Based ELISA

Assay Genie's GADD153 Colorimetric Cell-Based ELISA Kit is an in-cell ELISA designed for the quantitative detection of GADD153 protein levels in cells. It is intended as a lysate-free, high-throughput, sensitive assay for measuring the relative amounts of GADD153 in cultured cells and for screening the effects that various treatments, inhibitors (such as siRNA or chemicals) or activators have on GADD153.

Reactive against human, mouse and rat samples, the kit uses an indirect ELISA format in which GADD153 is captured by GADD153-specific primary antibodies and HRP-conjugated secondary antibodies bind the Fc region of the primary antibody, catalysing a colorimetric reaction on substrate addition that is read at 450 nm. The dynamic range is reported as > 5000 cells, and the kit is supplied in a microplate format for storage at 4 °C with a six-month shelf life. A monoclonal antibody specific for human GAPDH is included as an internal positive control for normalising target absorbance values, and a Crystal Violet whole-cell staining step may be used to determine cell density so that absorbance values can be normalised to cell amounts.

GADD153, also known as CHOP (C/EBP homologous protein), is a transcription factor involved in the cellular stress response and apoptosis, with described roles in ER stress, diabetes and neurodegenerative disease research. The associated gene symbol is DDIT3 (Gene ID 1649; UniProt ID P35638).