Description
Intended Uses: The Human high sensitivity IFNg ELISA Kit is to be used for the in-vitro quantitative determination of interferon gamma (IFNg) in human sera, plasmas, buffered solutions or cell culture media. The assay will recognize both natural and recombinant human IFNg.
Principle of the Assay: The IFNg Kit is a solid phase sandwich Enzyme Linked-Immuno- Sorbent Assay (ELISA). A monoclonal antibody specific for IFNg has been coated onto the wells of the microtiter strips provided. Samples, including standards of known IFNg concentrations and unknowns are pipetted into these wells. During the first incubation, the IFNg antigen and a biotinylated monoclonal antibody specific for IFNg are successively incubated. After washing, the enzyme (streptavidin-peroxydase) is added. All the unbound enzyme is removed by washing and the first amplification step is performed by adding the Biotine-Tyramine reagent. Under the action of HRP, a biotine polymerisation reaction occurs in the region of the HRP linked to the detection antibody. After washing the second amplification step is performed and the polymerised biotine is revealed by a new streptavidin-HRP step. Finally after washing, the substrate is added. The intensity of this coloured product is directly proportional to the concentration of IFNg present in the samples