Elk1 (Phospho-Ser389)Colorimetric Cell-Based ELISA Kit

A cell-based phospho-specific ELISA from Assay Genie designed for accurate detection of ELK1 phosphorylated at serine 389 in cell lysates. The qualitative indirect format captures the phospho-target with specific primary antibodies, while HRP-conjugated secondary antibodies bind the Fc region of the primary antibody and catalyse a colorimetric reaction at 450 nm upon substrate addition. A monoclonal antibody specific for human GAPDH is included as an internal positive control for normalising target absorbance, and Crystal Violet whole-cell staining may be used to determine cell density and adjust for plating differences. The kit offers high sensitivity and specificity with easy-to-follow protocols and quick assay times.

The lysate-free, high-throughput assay can be used to measure relative levels of phosphorylated Elk1 in cultured cells and to screen treatments, inhibitors (such as siRNA or chemicals) or activators for effects on Elk1 phosphorylation. Reactivity covers Human, Mouse and Rat samples, with a dynamic range greater than 5000 cells and detection by colorimetric measurement at 450 nm. Phosphorylation of ELK1 at serine 389 is mediated by MAP kinase pathways and is involved in cellular processes including cell growth, differentiation and survival; its dysregulation has been implicated in cancer, neurodegenerative disorders and inflammatory conditions. The product is classified under the Epigenetics and Nuclear Signaling research area.

The protocol begins by seeding adherent cells, fixing, quenching and blocking the plate, followed by overnight incubation at 4 °C with primary antibodies (anti-Elk1 Phospho-Ser389 and/or anti-Elk1 plus anti-GAPDH). After application of HRP-conjugated anti-rabbit or anti-mouse secondary antibodies, ready-to-use substrate is added and the reaction stopped before optical density is read at 450 nm.