IL-10 High Sensitivity Kit (Human)

Four sera and one cell culture medium samples with various concentrations of IL-10 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 4.3% and 6.3% respectively.

The protein encoded by this gene is a cytokine produced primarily by monocytes and to a lesser extent by lymphocytes. This cytokine has pleiotropic effects in immunoregulation and inflammation. It down-regulates the expression of Th1 cytokines, MHC class II Ags, and costimulatory molecules on macrophages. It also enhances B cell survival, proliferation, and antibody production. This cytokine can block NF-kappa B activity, and is involved in the regulation of the JAK-STAT signaling pathway. Knockout studies in mice suggested the function of this cytokine as an essential immunoregulator in the intestinal tract. Mutations in this gene are associated with an increased susceptibility to HIV-1 infection and rheumatoid arthritis.

Aviva Systems Biology IL-10 High Sensitivity ELISA Kit (Human) (OKDB00042) is designed for the highly sensitive in-vitro quantitative measurement of recombinant or natural IL-10 in human serum, plasma or cell culture media. 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-10 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-10 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 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 two concentrations of recombinant IL-10 in human serum and culture media in three experiments. Recoveries ranged from 70% to 101% with an overall mean recovery of 85%.

Ten specificities were tested with concentrations higher than IL-10 curve concentrations. No cross reaction was observed for concentrations for IL-1alpha and beta, IL-2, IL-5, IL-6, IL-8, IL-12, TNF-alpha, IFN-gamma and mIL-10.