IL-32? is the shortest and most abundant of four potential splice variants of the proinflammatory cytokine IL-32 (previously called NK4) with a predicted unmodified size of 15 kDa. Potential modifications include myristoylation and N-glycosylation. The IL-32? shows increased potency at inducing CXCL2/MIP-2 and CXCL8 expression in PBMC relative to uncleaved IL-32?. Transfected IL-32? was more likely to be cellassociated as compared to IL-32?, suggesting an intracellular function. IL-32 is induced by mitogens in peripheral lymphocytes, by IFN? in epithelial cells, or by IL-12 with IL-18 in NK cells and in turn induces cytokine expression.
For the quantitative determination of human interleukin 32?(IL-32?) concentrations in cell culture supernates, serum, and plasma.
Principle of the assay: This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for IL-32? has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any IL-32? present is bound by the immobilized antibody. Following incubation unbound samples are removed during a wash step, and then a detection antibody specific for IL-32? is added to the wells and binds to the combination of capture antibody- IL-32? in sample. Following a wash to remove any unbound combination, and enzyme conjugate is added to the wells. Following incubation and wash steps a substrate is added. A coloured product is formed in proportion to the amount of IL-32? present in the sample. The reaction is terminated by addition of acid and absorbance is measured at 450nm. A standard curve is prepared from seven IL-32? standard dilutions and IL-32? sample concentration determined.
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