IFN gamma High Sensitivity ELISA Kit (Human)

Two human serum pools, two human plasma pools and two cell culture media samples with various concentrations of IFN gamma were tested for repeatability and reproductibility. Each assay was carried out with 3 replicates of each sample. Three independent assays were performed. The intra-assay and inter-assay coefficient of variation has been calculated to be 3.9% and 8.6% respectively.

This gene encodes a soluble cytokine that is a member of the type II interferon class. The encoded protein is secreted by cells of both the innate and adaptive immune systems. The active protein is a homodimer that binds to the interferon gamma receptor which triggers a cellular response to viral and microbial infections. Mutations in this gene are associated with an increased susceptibility to viral, bacterial and parasitic infections and to several autoimmune diseases.

Aviva Systems Biology IFN gamma High Sensitivity ELISA Kit (Human) (OKDB00041) is designed for the highly sensitive in-vitro quantitative measurement of recombinant or natural IFN gamma 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 IFN gamma 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 IFN gamma 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 IFN gamma 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 levels of IFN gamma into three human serum pools, two plasma pools and two culture media. Recovery was evaluated with one test. Mean recoveries were 92% in sera (range values: 84-98%), 72% in plasma (Range values: 61-81%) and 108% in cell culture media (range values: 88-117%)

Ten specificities were tested with concentrations higher than IFNgamma curve concentrations. No cross reaction was observed for concentrations ranging from 2500 to 78.12 pg/ml for IL-a, IL-2, IL-8, IL-12p40, TNFa, CD95 Fas, TRAIL, ICAM-1, Gp130 and GM-CSF.