Interleukin-12 (IL12; formerly NKSF, for natural killer cell stimulatory factor, or CLMF, for cytotoxic lymphocyte maturation factor) is a novel cytokine cloned from B-cell lines. IL-12 is a heterodimeric molecule composed of p35 and p40 subunits (1). The larger 40kD subunit (p40) is a member of the cytokine receptor family, and the smaller 35kD subunit (p35) is related to IL-6 and GCSF (2). Both IL-12 p40(-/-) and p35(-/-) mice fail to produce IL-12 p70 heterodimer (3). Interleukin (IL)-12 has been cloned on the basis of its ability to activate natural killer (NK) cells and promote the development of cytolytic T cells. With further understanding of its activities, IL-12 has emerged as an important cytokine, affecting both immune and hematologic functions. It has been shown to be necessary for the T cell independent induction of interferon (IFN)-gamma, critical for the initial suppression of bacterial and parasitic infection; for the development of a Th1 response, critical for effective host defense against intracellular pathogens; and for the activation of differentiated T lymphocytes of both CD4+ and CD8+ phenotype (4). The subunits map to different chromosomes: p40 (IL-12B) to 5q31-q33 and p35 (IL-12A) to 3p12-3q13.2 (2). The standard product used in this kit is recombinant human IL-12(p40), which is a 40kD glycoprotein consisting of 306 amino acids.
Intended Use:IL-12 (p40) BioAssay™ ELISA Kit, Human is a sandwich ELISA for quantitative detection of human IL-12 (p40) in cell culture supernates, serum and plasma (heparin, EDTA, citrate).
Range:31.2-2000pg/ml
Sensitivity:< 2pg/ml
Specificity:Natural and recombinant human IL-12 (p40)
Crossreactivity:Crossreactivity with IL-12 (p70), IL-23 (p19/p40) < 5%
Assay Summary:1. Add samples and standards and incubate the plate at 37°C for 90 minutes. Do not wash. 2. Add biotinylated antibodies and incubate the plate at 37°C for 60 minutes. Wash plate 3 times with 0.01M TBS
3. Add ABC working solution and incubate the plate at 37°C for 30 minutes. Wash plate 5times with 0.01M TBS
4. Add TMB color developing agent and incubate the plate at 37°C in dark for 25-30 minutes
5. Add TMB stop solution and read..
Storage and Stability:Store *143891B powder at 4°C. Once reconstituted store at 4°C for up to 12 hours or at -20°C for up to 48 hours. Store other components at 4°C. Stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
