RFWD2 Colorimetric Cell-Based ELISA Kit

A cell-based assay from Assay Genie uses a colorimetric format to detect RFWD2 protein in cultured cell samples, providing a high-throughput, lysate-free workflow. The kit employs an indirect ELISA format in which RFWD2 is captured by RFWD2-specific primary antibodies, with HRP-conjugated secondary antibodies binding the Fc region of the primary antibody. Signal is generated by an HRP-catalyzed colorimetric reaction read at 450 nm, and the kit is intended for use with adherent cell samples. A monoclonal antibody specific for human GAPDH is included as an internal positive control for normalization, and Crystal Violet whole-cell staining may be used to determine cell density, allowing absorbance values to be normalized to cell amounts.

The assay shows reactivity with human and mouse samples, has a stated dynamic range of greater than 5000 cells, and is supplied in a 96-well microplate format. Recommended storage is at 4°C with a stability of 6 months. The kit is suitable for measuring relative RFWD2 levels in cultured cells and for screening the effects of treatments, inhibitors such as siRNA or chemicals, or activators on RFWD2 expression.

RFWD2, also known as COP1, is an E3 ubiquitin-protein ligase that mediates ubiquitination and subsequent proteasomal degradation of target proteins, including a documented role in p53 (TP53) ubiquitination and degradation and a function in the DNA damage response. The target is documented in the UniProt Knowledgebase under accession Q8NHY2, with Gene ID 64326 and OMIM entry 608067, and a stated molecular weight of 14,914 Da. The kit is listed under the Signal Transduction research area.