预期应用
ELISA法定量测定猪血清、血浆、细胞培养物上清或其它相关液体中IL-6含量。
实验原理
本试剂盒应用双抗体夹心酶标免疫分析法测定标本中白介素-6水平。用纯化的抗体包被微孔板，制成固相抗体，往包被单抗的微孔中依次加入白介素-6抗原、生物素化的抗猪白介素-6抗体、HRP标记的亲和素，经过彻底洗涤后用底物TMB显色。TMB在过氧化物酶的催化下转化成蓝色，并在酸的作用下转化成最终的黄色。颜色的深浅和样品中的白介素-6呈正相关。用酶标仪在450nm波长下测定吸光度（OD值），计算样品浓度。
Intended use
This immunoassay kit allows for the specific measurement of porcine IL-6 concentrations in cell culture supernates, serum, and plasma.
Introduction
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 various cells is regulated, either positively or negatively, by a variety of signals including mitogens, antigenic stimulation, lipopolysaccharides, IL-1, TNF, PDGF and viruses. On the basis of its various activities, IL-6 has also been called interferon-β2 (IFN-β2), 26 kDa protein, B-cell stimulatory factor-2 (BSF-2), hybridoma/plasmacytoma growth factor, hepatocyte stimulating factor, cytotoxic T-cell differentiation factor, and macrophage-granulocyte inducing factor 2A (MGI-2A).
The IL-6 cDNA sequence predicts a protein of 212 amino acid residues in length with two potential N-glycosylation sites. The hydrophobic N-terminal 28 amino acid 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. Mouse IL-6 cDNA sequence shows a homology of 42% at the amino acid level when compared with the human sequence. Sequencing of the genomic DNA for IL-6 indicates that the gene for this factor consists of five exons and four introns. 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.
Test principle
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. An enzyme-linked monoclonal antibody specific for IL-6 is added to the wells. Following a wash to remove any unbound antibody-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of IL-6 bound in the initial step. The color development is stopped and the intensity of the color is measured. |
