IRF-3 (Phospho-Ser396)Colorimetric Cell-Based ELISA Kit

From Assay Genie, this cell-based ELISA kit is designed to detect and quantify phosphorylated IRF-3 at Ser396 in cultured cells, with reactivity confirmed against human, mouse, and rat samples. The kit employs a colorimetric detection method measured at 450 nm, and is validated for a dynamic range of more than 5,000 cells.

The assay uses an indirect ELISA format in which IRF-3 (Phospho-Ser396) is captured by target-specific primary antibodies and detected with HRP-conjugated secondary antibodies that catalyze a colorimetric reaction upon substrate addition. A monoclonal antibody specific for human GAPDH is included as an internal positive control to normalize target absorbance values, and a Crystal Violet whole-cell staining method may be used to determine cell density and adjust for plating differences. The phospho-specific design makes the kit suitable for screening the effects of various treatments, inhibitors, or activators on IRF-3 phosphorylation.

IRF-3 is a DNA-binding transcription factor of the interferon regulatory factor family that resides in the cytoplasm in an inactive form. Phosphorylation at Ser396 is a key activation event that enables IRF-3 to form a complex with CREBBP, translocate to the nucleus, and drive transcription of interferons alpha and beta together with other interferon-induced genes involved in the antiviral defense response.