ASK1 (Phospho-Ser83)Colorimetric Cell-Based ELISA Kit

This Colorimetric Cell-Based ELISA Kit from Assay Genie is designed for the accurate detection of phosphorylated serine 83 levels on ASK1 in cell lysates and cell culture samples. The assay is configured for human samples and employs a lysate-free, high-throughput indirect ELISA workflow in which ASK1 (Phospho-Ser83) is captured by target-specific primary antibodies, followed by HRP-conjugated secondary antibodies that catalyze a colorimetric reaction measured at 450 nm. Qualitative readouts of phosphorylated ASK1 are supported by two normalization strategies: a monoclonal anti-human GAPDH antibody supplied as an internal positive control, and a Crystal Violet whole-cell staining step to account for cell density differences across wells.

The dynamic range is greater than 5000 cells, and the kit is suitable for measuring the relative amounts of phosphorylated ASK1 in cultured adherent cells and for screening the effects of treatments, inhibitors (such as siRNA or chemical compounds), or activators on ASK1 phosphorylation. The target corresponds to MAP3K5 (Gene ID 4217, UniProt ID Q99683, OMIM 602448), a MAP kinase kinase kinase of the STE11 family also known as apoptosis signal-regulating kinase 1, ASK-1, MEKK5, or MAPKKK5, with a molecular weight of 69,839 Da. ASK1 is activated by stressors such as oxidative stress, endoplasmic reticulum stress, and calcium overload, as well as by inflammatory signals including TNF and LPS, and it phosphorylates the downstream MAP kinase kinases MKK4/SEK1 and MKK3/MKK6 to activate JNK and p38 MAPK pathways, linking it to stress-induced apoptosis and innate immune responses.