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Mouse IL-6 Quantikine ELISA Kit Summary

Product Summary

The Quantikine Mouse IL-6 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure mouse IL-6 in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant mouse IL-6 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant mouse IL-6. Results obtained using natural mouse IL-6 showed dose response curves that were parallel to the standard curves obtained using the Quantikine mouse kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse IL-6.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision

Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision

Cell Culture Supernates, Serum, Heparin Plasma

Recovery

The recovery of mouse IL-6 spiked to three levels throughout the range of the assay in various matrices was evaluated.

Stability and Storage

Supporting data and figures

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of mouse IL-6 were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.

Mouse IL-6 ELISA Standard Curve

Background

IL-6

Interleukin 6 (IL-6) is a pleiotropic, a-helical, 22-28 kDa phosphorylated and variably glycosylated cytokine that plays important roles in the acute phase reaction, inflammation, hematopoiesis, bone metabolism, and cancer progression. Mature human IL-6 is 183 amino acids (aa) in length and shares 39% aa sequence identity with mouse and rat IL-6. Alternative splicing generates several isoforms with internal deletions, some of which exhibit antagonistic properties. Cells known to express IL-6 include CD8+ T cells, fibroblasts, synoviocytes, adipocytes, osteoblasts, megakaryocytes, endothelial cells (under the influence of endothelins), sympathetic neurons, cerebral cortex neurons, adrenal medulla chromaffin cells, retinal pigment cells, mast cells, keratinocytes, Langerhans cells, fetal and astrocytes, neutrophils, monocytes, eosinophils, colonic epithelial cells, B1 B cells and pancreatic islet beta cells. IL-6 production is generally correlated with cell activation and is normally kept in control by glucocorticoids, catecholamines, and secondary sex steroids. Normal human circulating IL-6 is in the 1 pg/mL range, with slight elevations during the menstrual cycle, modest elevations in certain cancers, and large elevations after surgery.

IL-6 induces signaling through a cell surface heterodimeric receptor complex composed of a ligａnd binding subunit (IL-6 R alpha) and a signal transducing subunit (gp130). IL-6 binds to IL-6 Ra, triggering IL-6 Ra association with gp130 and gp130 dimerization. gp130 is also a component of the receptors for CLC, CNTF, CT-1, IL-11, IL-27, LIF, and OSM. Soluble forms of IL-6 Ra are generated by both alternative splicing and proteolytic cleavage. In a mechanism known as trans-signaling, complexes of soluble IL-6 and IL-6 Ra elicit responses from gp130-expressing cells that lack cell surface IL-6 Ra. Trans-signaling enables a wider range of cell types to respond to IL-6, as the expression of gp130 is ubiquitous, while that of IL-6 Ra is predominantly restricted to hepatocytes, monocytes, and resting lymphocytes. Soluble splice forms of gp130 block trans-signaling from IL-6/IL-6 Ra but not from other cytokines that use gp130 as a co-receptor.

IL-6, along with TNF-a and IL-1, drives the acute inflammatory response. IL-6 is almost solely responsible for fever and the acute phase response in the liver, and it is important in the transition from acute inflammation to either acquired immunity or chronic inflammatory disease. When dysregulated, it contributes to chronic inflammation in conditions such as obesity, insulin resistance, inflammatory bowel disease, arthritis, and sepsis. IL-6 modulates bone resorption and is a major effector of inflammatory joint destruction in rheumatoid arthritis through its promotion of Th17 cell development and activity. It contributes to atherosclerotic plaque development and destabilization as well as the development of inflammation-associated carcinogenesis.