A-RAF Colorimetric Cell-Based ELISA Kit

From Assay Genie, the A-RAF Colorimetric Cell-Based ELISA Kit is a lysate-free, high-throughput in-cell assay designed to detect A-RAF protein expression profiles directly in cultured cells. The kit uses an indirect ELISA format in which A-RAF is captured by A-RAF-specific primary antibodies and detected via HRP-conjugated secondary antibodies that catalyze a colorimetric reaction upon substrate addition, with absorbance read at 450 nm. It is suited for measuring relative A-RAF levels in cultured cells and for screening the effects of treatments, inhibitors (e.g. siRNA or chemicals), or activators on A-RAF expression. Sample type is adherent cells.

A monoclonal antibody specific for human GAPDH is included as an internal positive control for normalising target absorbance values. Following the colorimetric HRP measurement, an optional Crystal Violet whole-cell staining step may be used to determine cell density, allowing absorbance values to be normalised to cell amounts and correct for plating variation. The kit has been validated for reactivity with Human, Mouse and Rat samples and shows a dynamic range of > 5000 Cells.

The assay supports research into A-RAF (ARAF; UniProt P10398; NCBI Gene ID 369), a serine/threonine kinase of the RAF family implicated in mitogenic signalling, cell growth and differentiation, and dysregulated in cancers such as thyroid cancer, melanoma and colorectal cancer. Storage is at 4°C for 6 months.