B-RAF (Phospho-Ser446)Colorimetric Cell-Based ELISA Kit

From Assay Genie, this Cell-Based ELISA kit is designed to detect phosphorylation of B-RAF at Ser446 in cultured cells, providing a lysate-free, high-throughput workflow for studying cell signalling and kinase activity. The assay uses an indirect ELISA format in which B-RAF (Phospho-Ser446)-specific primary antibodies capture the phosphorylated target, while HRP-conjugated secondary antibodies bind the primary antibody Fc region and catalyse a colorimetric reaction upon substrate addition, enabling qualitative measurement of relative B-RAF phosphorylation under different treatment, inhibitor or activator conditions.

Detection is performed by absorbance measurement at 450 nm, with a stated dynamic range of > 5000 Cells. The kit is reactive with Human and Mouse samples and is intended for adherent cells. A monoclonal anti-GAPDH antibody is included as an internal positive control for normalising target absorbance values, and Crystal Violet whole-cell staining is supplied so that cell density can be determined and readings adjusted for plating differences.

The target corresponds to the BRAF gene product (Gene ID: 673; UniProt ID: P15056), a serine/threonine kinase of the RAF family with a reported molecular weight of 84 kDa. The sub-type is Phospho, and the kit is positioned for research applications including cancer biology, drug development and MAPK signalling pathway studies, where monitoring Ser446 phosphorylation can inform understanding of B-Raf activation dynamics and potential therapeutic targets such as Nexavar/Sorafenib.