SEK1/MKK4 (Phospho-Thr261)Colorimetric Cell-Based ELISA Kit

A cell-based ELISA kit from Assay Genie for the qualitative determination of SEK1/MKK4 phosphorylated at threonine 261. The assay is lysate-free and uses an indirect ELISA format in which the phospho-target is captured by target-specific primary antibodies, while HRP-conjugated secondary antibodies bind the Fc region of the primary antibody and catalyze a colorimetric reaction upon substrate addition. It is intended for measuring the relative amounts of phosphorylated SEK1/MKK4 in cultured cells and for screening the effects that various treatments, inhibitors (such as siRNA or chemicals), or activators have on SEK1/MKK4 phosphorylation.

The kit is reactive with Human, Mouse and Rat samples, with a stated dynamic range of > 5000 Cells and colorimetric detection read at 450 nm. Sample type is adherent cells. Because the Cell-Based ELISA is qualitative, two normalization strategies are included: a monoclonal antibody specific for human GAPDH serves as an internal positive control for normalizing target absorbance values, and a Crystal Violet whole-cell staining step may be performed after HRP measurement to determine cell density, allowing absorbance values to be normalized to cell counts.

SEK1/MKK4 (UniProt ID: P45985; Gene ID: 6416; Gene Symbol: MAP2K4) is a dual-specificity kinase of the STE7 family that phosphorylates and activates JNK1 and JNK2 as well as p38, but not ERK1 or ERK2, and mediates cellular responses to various cellular stresses and inflammatory cytokines. Phosphorylation of SEK1/MKK4 at threonine 261 is described as regulating its activity, making the phospho-specific kit a tool for studying MAPK signaling in cell growth, proliferation and survival.