C-RAF (Phospho-Ser296)Colorimetric Cell-Based ELISA Kit

Assay Genie's Colorimetric Cell-Based ELISA Kit is designed for the precise measurement of C-RAF phosphorylation at Serine 296 in cell lysates. As a Cell-Based (In-Cell) ELISA, the assay is lysate-free and supports high-throughput workflows, enabling researchers to measure relative levels of phosphorylated C-RAF in cultured cells and to screen the effects of treatments, inhibitors (such as siRNA or chemicals), and activators on C-RAF phosphorylation.

Detection is achieved by an indirect ELISA format: C-RAF (Phospho-Ser296) is captured by a phospho-specific primary antibody, and an HRP-conjugated secondary antibody binds the Fc region of that primary antibody, catalysing a colorimetric reaction upon substrate addition at 450 nm. The kit shows reactivity with Human, Mouse and Rat samples and has a dynamic range of > 5000 Cells, with the supplied microplates suited to adherent cell culture. For normalisation, a monoclonal anti-GAPDH antibody is included as an internal positive control, and Crystal Violet whole-cell staining can be used to determine cell density and adjust for plating differences.

C-RAF is a key regulator of cell signalling, controlling growth, differentiation and survival, and phosphorylation at Serine 296 is a critical event in its activation. Dysregulation of this modification is implicated in diseases such as cancer and inflammatory disorders, making this kit well suited to studies of cell signalling and to the development of targeted therapies.