MADD Colorimetric Cell-Based ELISA

An in-cell ELISA kit from Assay Genie is designed for the qualitative detection of MADD (MAP-kinase activating death domain protein) in cultured cells. Using an indirect ELISA format, the assay captures MADD with target-specific primary antibodies and detects it with HRP-conjugated secondary antibodies that catalyze a colorimetric reaction measured at 450 nm. The kit is reactive with human, mouse, and rat samples and is intended for adherent cells.

Researchers can use it to measure the relative amounts of MADD in cultured cells and to screen for the effects of treatments, inhibitors such as siRNA or chemicals, and activators. Because the assay is qualitative, two normalisation methods are provided: an anti-GAPDH monoclonal antibody as an internal positive control, and a Crystal Violet whole-cell staining step to determine cell density, which allows absorbance values to be adjusted for plating differences. The stated dynamic range is > 5000 Cells, the format is a 96-Well Microplate, and the kit is intended for storage at 4 °C with a 6-month stability.

MADD is a guanine nucleotide exchange factor that activates RAB3A/C/D and Rab27A/B, and it plays a significant role in cell signalling, proliferation, and apoptosis regulation. Dysregulation of MADD has been linked to cancer and neurodegenerative disorders, making it a useful target for cell biology, molecular biology, and drug discovery research.