IL-6 High Sensitivity ELISA Kit (Human)

Four sera and one cell culture medium samples with various concentrations of IL-6 were tested for repeatability and reproducibility. Each assay was carried out with 3 replicates of each sample. Five independent assays were performed. The intra-assay and inter-assay coefficient of variation has been calculated to be 4.4% and 9.1% respectively.

This gene encodes a cytokine that functions in inflammation and the maturation of B cells. In addition, the encoded protein has been shown to be an endogenous pyrogen capable of inducing fever in people with autoimmune diseases or infections. The protein is primarily produced at sites of acute and chronic inflammation, where it is secreted into the serum and induces a transcriptional inflammatory response through interleukin 6 receptor, alpha. The functioning of this gene is implicated in a wide variety of inflammation-associated disease states, including suspectibility to diabetes mellitus and systemic juvenile rheumatoid arthritis. Alternative splicing results in multiple transcript variants.

Aviva Systems Biology IL-6 High Sensitivity ELISA Kit (Human) (OKDB00046) is designed for the highly sensitive in-vitro quantitative measurement of recombinant or natural IL-2 in supernatants, buffered solutions or serum and plasma samples and other body fluids. The kit is based on standard sandwich enzyme-linked immuno-sorbent assay methods along with an additional detection process employing a polymerization step to amplify the number of detector bound HRP molecules. A monoclonal antibody specific for IL-6 has been pre-coated into wells of a 96-well plate. Standards or test samples are added to the wells and incubated. After washing, a biotinylated detector antibody specific for IL-6 is added, incubated and followed by washing. Streptavidin-HRP conjugate is then added, incubated and unbound conjugate is washed away. Amplification is achieved by adding a Biotin-Tyramine reagent, incubating and washing, followed by a second incubation with Streptavidin-HRP conjugate. An enzymatic reaction is then produced through the addition of TMB substrate which is catalyzed by HRP to generating a blue color product that changes yellow after adding acidic stop solution. The density of yellow coloration read by absorbance at 450 nm and is quantitatively proportional to the amount of sample IL-6 captured in well.

Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Reconstitution & Storage Instructions Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Antibody Array (AA) Protocol

The spike recovery was evaluated by spiking 3 concentrations of IL-6 into human serum in 2 separate experiments. Recoveries ranged from 107% to 123% with an overall mean recovery of 115%.

The assay recognizes both natural and recombinant human IL-6. To define the specificity of this ELISA several proteins were tested for cross reactivity. There was no cross reactivity observed for any protein tested (IL-1a, IL-b, IL-10, IL-12, IFNgamma, IL-4, TNFa, IL-8 and IL-13) .