human IL-6 ELISA kit (48 Wells)

Interleukin 6(IL-6) is a multifunctional protein produced by lymphoid and non-lymphoid cells and by normal and transformed cells, including T cells, monocyte/macrophages, fibroblasts, hepatocytes, vascular endothelial cells, cardiac myxomas, bladder cell carcinomas, myelomas, astrogliomas and glioblastomas. The production of IL-6 in these cells is regulated, either positively or negatively, by a variety of signals including mitogens, antigenic stimulation, lipopolysaccharides, IL-1, TNF, PDGF and viruses. For reviews on IL-6, see references 1-5.
The human IL-6 cDNA sequence predicts a protein of 212 amino acid (aa) residues in length with two potential N-glycosylation sites. The hydrophobic N-terminal 28 aa residue signal peptide is cleaved to produce a mature protein of 184 amino acids with four cysteine residues and a predicted molecular mass of 21 kDa (6-9). The mouse IL-6 cDNA sequence shows a homology of 42% at the aa level when compared with the human sequence (10). On the basis of sequence similarity and gene structural motif similarity, IL-6 can be grouped in a family of cytokines that also includes OSM, G-CSF, LIF, and CNTF. All of these cytokines are predicted to have a four helix bundle structure similar to that found for growth hormone, suggesting that they all evolved from a common ancestral gene (11-13).
The effects of IL-6 on different cells are numerous and varied. The effect on B cells is stimulation of differentiation and antibody secretion (6, 14 - 17). IL-6 also affects T cells, acting as a co-stimulant with sub-optimal concentrations of PHA or Con A to stimulate IL-2 production and IL-2 receptor expression.
IL-6 exhibits growth factor activity for mature thymic or peripheral T-cells and reportedly enhances the differentiation of cytotoxic T-cells in the presence of IL-2 or IFN-? (18-20). IL-6 stimulates production of acute phase proteins by hepatocytes (21) and has colony-stimulating activity on hematopoietic stem cells. IL-6 has growth factor activities and will stimulate the growth of myeloma/hybridoma/ plasmacytoma cells, EBV-transformed B cells, keratinocytes and mesangial cells (4, 5).

For the quantitative determination of human interleukin 6 (IL-6) 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-6 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any IL-6 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-6 is added to the wells and binds to the combination of capture antibody-IL-6 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-6 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-6 standard dilutions and IL-6 sample concentration determined.

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