MAP2K7 (Phospho-Thr275)Colorimetric Cell-Based ELISA Kit

This cell-based ELISA kit from Assay Genie is designed for the accurate detection of phosphorylated MAP2K7 at Thr275 in cell lysates and tissue homogenates. MAP2K7, also known as MEK7, is a dual-specificity protein kinase within the MAPK signalling pathway that regulates cell growth, differentiation and stress responses. Phosphorylation at Thr275 plays a crucial role in mediating its activity and downstream signalling cascades, and dysregulation of this pathway has been implicated in cancer, inflammation and neurodegenerative disorders.

The assay uses an indirect ELISA format, capturing the phospho-target with specific primary antibodies while HRP-conjugated secondary antibodies bind the Fc region of the primary antibody, enabling a colorimetric reaction at 450 nm. It is a lysate-free, high-throughput assay suitable for measuring relative levels of phosphorylated MAP2K7 in cultured cells and for screening the effects of various treatments, inhibitors such as siRNA or chemicals, and activators on MAP2K7 phosphorylation. The kit shows reactivity with Human, Mouse and Rat samples and has a dynamic range of > 5000 Cells.

Two normalisation methods are provided: a monoclonal anti-GAPDH antibody serves as an internal positive control for normalising target absorbance values, while Crystal Violet whole-cell staining may be used to determine cell density so that absorbance values can be adjusted to cell amounts. The procedure involves seeding adherent cells, fixing, quenching, blocking, overnight primary antibody incubation at 4°C, secondary HRP-conjugated antibody incubation, substrate addition and a stop step, with washes between stages.