Capture Antibody: Pre-titrated, purified antibody Detection Antibody: Pre-titrated, biotin-conjugated antibody Standard: Recombinant protein for generating standard curve and calibrating samples 10X Coating Buffer: Buffer for plating the Capture Antibody 5X ELISA/ELISPOT Diluent: Buffer for blocking and diluting the Detection Antibody and Enzyme Enzyme: Pre-titrated Avidin-HRP Substrate: 1X TMB Solution Certificate of Analysis: Lot-specific instructions for dilution of antibodies and standards 96 Well Plate: Corning Costar 9018 (included with product Cat. Nos. ending in suffixes -22, -76, -86)
The Human Interferon gamma (IFNγ) Uncoated ELISA Kit contains pre-matched antibody pairs, plates and reagents for performing quantitative enzyme linked immunosorbent assays (ELISA) to detect and quantify protein levels of human IFNγ. Wash Buffer and Stop Solution are needed to complete the ELISA reaction and are sold separately.
Principle of the method
ELISAs are designed to measure the amount of the target bound between a matched antibody pair. A target-specific antibody is coated to the bottom of the wells of a microplate, which is an overnight process. Samples, standards, or controls are then added into these wells and bind to the immobilized (capture) antibody. A sandwich is formed by the addition of the second (detector) antibody, a substrate solution is added that reacts with the enzyme-antibody-target complex to produce measurable signal. The intensity of this signal is directly proportional to the concentration of target present in the original specimen.
IFN gamma (Interferon gamma, Type II interferon) is a macrophage activation factor, and immune interferon that is produced primarily by T-lymphocytes and natural killer cells in response to antigens, mitogens, Staphylococcus enterotoxin B, phytohemaglutanin and other cytokines. IFN gamma is a dimeric protein consisting of two 146 amino acid subunits. IFN gamma is a glycoprotein that exists, functionally, as a homodimer of approximately 45 kDa. On SDS-PAGE, IFN gamma appears as a combination of 25, 20 and minor 15.5 kDa bands as a result of differential glycosylation. The biological activity of the IFN gamma homodimer is highly species specific. Human IFN gamma does not show cross-reactivity with mouse. IFN gamma function includes the following: antiviral activity, tumor antiproliferative activity, induction of class I and II MHC, macrophage activation, and enhanced immunoglobulin secretion by B lymphocytes. IFN gamma is involved in cytokine regulation and also acts synergistically with other cytokines. Activation of IFN gamma takes place through binding of IFN gamma receptor I and II, and activating the JAK-STAT pathway. IFN gamma does not show any homology with IFN alpha or IFN beta but human IFN gamma shows about 40% sequence homology with mouse IFN gamma. IFN gamma is upregulated by IL2, FGF basic, EGF and downregulated by vitamin D3 or DMN. IFN gamma gene mutations are associated with aplastic anemia.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.