JNK1&2 ELISAs are designated for quantitative determination of human and mouse phosphorylated or non-phosphorylated JNK1&2 in cell lysates. The antibodies in the JNK1&2 ELISAs require that the protein in cells be denatured in order to achieve recognition. The assay is a solid-phase sandwich enzyme-linked immunosorbent assay (ELISA). A monoclonal capture antibody specific for the JNK1&2 has been coated onto the multiwell strips provided with the kit. Standard dilutions and samples are incubated for 2 hours at room temperature. JNK1&2 antigen binds to the capture antibody. After a wash, a detection antibody specific for the non-phosphorylated or phosphorylated JNK1&2 protein is incubated for 1 hr at room temperature, which results in binding of the antibody to the immobilized JNK1&2 protein. An anti-rabbit IgG-HRP binds to the immobilized protein, completing the four-member sandwich. The reaction is visualized by tetramethylbenzidine (TMB) substrate, followed by the stop solution. The intensity of the yellow color, measured in a multiwell plate reader at 450 nm, is directly proportional to the concentration of JNK1&2 in the original sample. The unknown concentrations are calculated from the standard curve run with each assay.
JNK plays a role in apoptosis and cell survival. JNK1 and JNK2 are required for polarized differentiation of T-helper cells into Th1 cells. Activation of JNK1&2 causes phosphorylation of a number of transcription factors, such as c-Jun and ATF2, and thus regulates AP-1 transcriptional activity.
Features and Benefits
JNK1&2 ELISAs are fast (total time 4 hours), sensitive (detects <0.15 ng/mL or <0.8 units/mL), and specific for either non-phosphorylated or phosphorylated JNK1&2. Precoated plates are provided. All incubations are at room temperature and no proprietary equipment is required.
JNK1&2 ELISA is specific for native and recombinant human and mouse JNK1&2 regardless of phosphorylation state. The sensitivity is <0.15 ng/mL and equals the sensitivity of immunobloting. Linear regression yielded a correlation coefficient of 0.99.