Principle of Assay
This IRF-3 (phospho Ser396) Cell Based ELISA Kit allows for the detection of IRF-3 (phospho Ser396) and the effects that certain stimulation conditions have on IRF-3 (phospho Ser396) expression in different cell lines. Qualitative determination of IRF-3 (phospho Ser396) concentration is achieved by an indirect ELISA format. In essence, IRF-3 (phospho Ser396) is captured by Anti-IRF-3 (phospho Ser396) Antibodies while Dye 1-conjugated and Dye 2-conjugated secondary antibodies bind the Fc region of the primary antibody. Through this binding, the dye conjugated to the secondary antibody can emit light at a certain wavelength given proper excitation, hence allowing for a fluorometric detection method. A monoclonal antibody specific for human GAPDH is included to serve as an internal positive control in normalizing the target RFU values. Anti-IRF-3 Antibody is also included for normalization purposes. The RFU values obtained for the non-phosphorylated target can be used to normalize the RFU value for the phosphorylated target.
