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EK000022-EK0421: Human IL-12 (p70) ELISA Kit
Range 7.8pg/ml-500pg/ml
Sensitivity < 2 pg/ml
Specificity: no detectable cross-reactivity with any other cytokine.
Application: for quantitive detection of human IL-12(p70) in sera, plasma, body fluids, tissue lysates or cell culture supernates.
Expiration: four months at 4°C and eight months at -20°C.
Background
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. The larger 40-kDa subunit (p40) is a member of the cytokine receptor family, and the smaller 35-kDa subunit (p35) is related to IL6 and GCSF. Both IL-12 p40(-/-) and p35(-/-) mice fail to produce IL-12 p70 heterodimer. 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. The subunits map to different chromosomes: p40 (IL12B) to 5q31-q33 and p35 (IL12A) to 3p12-3q13.2.2 The standard product used in this kit is recombinant human IL-12(p40), which is a 40KDa glycoprotein consisting of 306 amino acids.
Principle
The ELISA Kit is based on standard sandwich enzyme-linked immune-sorbent assay technology. The target specific antibodies are precoated onto 96-well plates. The target from the sample is bound to the microwell. The biotinylated target specific detection antibodies are added to the microwells and followed by washing with the PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex is added and unbound conjugates are washed away with the PBS or TBS buffer. TMB, the HRP (horseradish peroxidase) substrate, is used to visualize color change resulting from the HRP enzymatic reaction. TMB is catalyzed by HRP to produce the blue color. The color changes into yellow after the acidic stop solution is added. The density of the yellow color is proportional to the target amount from the sample captured in the microwells.
Reference
1. Toubai T, Tanaka J, Ota S, Fukuhara T, Hashino S, Kondo T, Shono Y, Morioka M, Kawamura T, Masauzi N, Kakinoki Y, Kobayashi H, Kunieda Y, Kasai M, Kurosawa M, Asaka M, Imamura M. Effect of granulocyte colony-stimulating factor on IL-12 p40 production during chemotherapy for B-cell lineage non-Hodgkin's lymphoma patients. Eur J Haematol. Nov;77(5):403-9.2006.
2. Nakamura T, Kimura H, Kato M, Kurashige S, Wakamatsu K. A sensitive and reliable quantification method for mouse interleukin-12 p70 based on fluorometric sandwich ELISA (FS-ELISA).Cell Biol Int. Feb;31(2):173-9.2007
3. Spensieri F, Fedele G, Fazio C, Nasso M, Stefanelli P, Mastrantonio P, Ausiello CM. Bordetella pertussis inhibition of interleukin-12 (IL-12) p70 in human monocyte-derived dendritic cells blocks IL-12 p35 through adenylate cyclase toxin-dependent cyclic AMP induction. Infect Immun. May;74(5):2831-8,2006.
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