B-Raf (Phospho-Thr753)Colorimetric Cell-Based ELISA Kit

From Assay Genie, this cell-based ELISA kit is designed for the qualitative measurement of B-Raf phosphorylation at threonine 753 in cultured cells using a lysate-free workflow. The indirect ELISA format captures B-Raf (Phospho-Thr753) with phospho-specific primary antibodies, while HRP-conjugated secondary antibodies catalyze a colorimetric reaction read at 450 nm. The kit is reactive with human, mouse, and rat samples, including adherent cells, and is suitable for screening the effects of treatments, siRNA, chemical inhibitors, or activators on B-Raf phosphorylation.

Because the assay is qualitative, two normalization methods are supplied: a monoclonal anti-GAPDH antibody acting as an internal positive control, and Crystal Violet whole-cell staining to assess cell density so absorbance values can be normalized to cell number. The dynamic range exceeds 5,000 cells. The target, B-Raf (gene symbol BRAF1), is a serine/threonine kinase of the RAF family that transduces mitogenic signals through the MAP kinase/ERKs pathway, and its dysregulation is implicated in thyroid cancers, skin melanomas, colorectal cancers, lung cancers, gliomas, and developmental disorders such as cardiofaciocutaneous syndrome.