Documents

CS0130

Sigma-Aldrich

phospho-JNK1&2 (pThr183/pTyr185) ELISA

sufficient for 96 tests

Synonym(s):
JNK 1/2 ELISA, c-Jun N-terminal Kinase (JNK)1&2 ELISA
Pricing and availability is not currently available.

usage

sufficient for 96 tests

shipped in

wet ice

storage temp.

2-8°C

Gene Information

human ... MAPK8(5599), MAPK9(5601)

General description

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.

Biochem/physiol Actions

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.

Analysis Note

phospho-JNK1&2 (pThr183/pTyr185) ELISA is specific for JNK1&2 dually phosphorylated at threonine 183 and tyrosine 185. The sensitivity is <0.8 units/mL, which is twice as sensitive as immunoblotting. Linear regression yielded a correlation coefficient of 0.99.

RIDADR

UN 3316 9/PG 2

Certificate of Analysis
Certificate of Origin
Signal transduction. MAP kinase signaling specificity.
Claire R Weston et al.
Science (New York, N.Y.), 296(5577), 2345-2347 (2002-06-29)

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