Interleukin-6 (IL-6) is a protein that in humans is encoded by the IL6 gene.1 Interleukin-6 (IL-6) is involved not only in the hepatic acute phase response but also in adipose tissue metabolism, lipoprotein lipase activity, and hepatic triglyceride secretion.2 IL-6 is a pleiotropic cytokine that plays a critical role in bone resorption.3 IL-6 has been suggested to stimulate the HPA axis during immune activation independent of the input of hypothalamic corticotropin-releasing hormone (CRH).4 Interleukin-6 (IL-6) is an immunoregulatory cytokine that activates a cell-surface signaling assembly composed of IL-6, the IL-6 alpha-receptor (IL-6Ralpha), and the shared signaling receptor gp130.5 The standard product used in this kit is recombinant rat IL-6, consisting of 188 amino acids with the molecular mass of 21.8kD. Intended Use: IL-6 BioAssay(TM) ELISA Kit is a sandwich ELISA for quantitative detection of rat IL-6 in cell culture supernates, serum and plasma (heparin, EDTA, citrate). Range: 62.5-4000pg/ml Sensitivity: <5pg/ml Specificity: Natural and recombinant rat IL-6 Crossreactivity: No detectable crossreactivity with other relevant proteins Kit Components: 143931A: Microtiter Strips, 1x96 wells *143931B: IL-6 Standard, 2x1 vial 143931C: Anti-rat IL-6 (Biotin), 1x130ul 143696: Avidin-Biotin-Peroxidase Complex (ABC), 1x130ul (1:100 dilution) 143697: Sample Diluent Buffer, 1x30ml 143698: Antibody Diluent Buffer, 1x12ml 143699: ABC Diluent Buffer, 1x12ml 143700: TMB Color Developing Agent, 1x10ml 143701: TMB Stop Solution, 2N H2SO4, 1x10ml Storage and Stability: Store *143931B powder at 4C. Once reconstituted store at 4C for up to 12 hours or at -20C for up to 48 hours. Store other components at 4C. Stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Assay Summary: 1. Add samples and standards and incubate the plate at 37C for 90 minutes. Do not wash. 2. Add biotinylated antibodies and incubate the plate at 37C for 60 minutes. Wash plate 3 times with 0.01M TBS or PBS. 3. Add ABC working solution and incubate the plate at 37C for 30 minutes. Wash plate 5 times with 0.01M TBS or PBS. 4. Add TMB color developing agent and incubate the plate at 37C in dark for 20-25 minutes. 5. Add TMB stop solution and read.
