Principle of the Assay
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for IFN-γ has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any IFN-γ present is bound by the immobilized antibody. Following incubation unbound samples are removed during a wash step, and then a detection antibody specific for IFN-γ is added to the wells and binds to the combination of capture antibody- IFN-γ in sample. Following a wash to remove any unbound combination, and enzyme conjugate is added to the wells. Following incubation and wash steps a substrate is added. A coloured product is formed in proportion to the amount of IFN-γ present in the sample. The reaction is terminated by addition of acid and absorbance is measured at 450nm. A standard curve is prepared from seven IFN-γ standard dilutions and IFN-γ sample concentration determined.
For Use with serum, plasma and cell culture supernatants. For Research Use Only. Not for use in diagnostic procedures.
Interferon gamma (IFN-γ) is a multifunctional protein first observed as an antiviral activity in cultures of Sindbis virus-infected human leukocytes stimulated by PHA. Produced by Tlymphocytes and natural killer (NK) cells, IFN-γ is now known to be both an inhibitor of viral replication and a regulator of numerous immunological functions. Human IFN-γ is reported to be active only on human and non-human primate cells. The biochemistry and biological activities of interferons have been extensively reviewed.
Human IFN-γ is a 143 amino acid residue, 20 or 25 kDa glycoprotein that demonstrates little sequence homology to IFN-α or -β. Naturally occurring IFN-γ is found as either of two molecular weight species, differing in degree of glycosylation. Human IFN-γ apparently exists as a head-to-tail dimer in solution with the C-terminus of one monomer aligned with the N-terminus of the other monomer A receptor for IFN-γ has been identified and its gene localized to chromosome 6 Apparently the product of a single gene, the receptor is a single chain 90 kDa glycoprotein that shows a high degree of species-specific binding of IFN-γ.
Functionally, IFN-γ produces a variety of effects. Produced by CD8+, NK, gd, and TH1 T helper cells, IFN-γ has documented antiviral, antiprotozoal and immunomodulatory effects on cell proliferation and apoptosis, as well as the stimulation and repression of a variety of genes he antiprotozoal activity of IFN-γ against Toxoplasma and Chlamydia is believed to result from indoleamine 2,3-dioxygenase activity, an enzyme induced by IFN-γ .The immunomodulatory effects of IFN-γ are extensive and diverse. In monocyte/macrophages, the activities of IFN-γ include: increasing the expression of class I and II MHC antigens; increasing the production of IL-1, platelet-activating factor, H2O2, and pterin; protection of monocytes against LAK cell-mediated lysis; downregulation of IL-8 mRNA expression that is upregulated by IL-2; and, with lipopolysaccharide, induction of NO production.Finally, IFN-γ has been shown to upregulate ICAM-1, but not E-Selectin or VCAM-1, expression on endothelial cells.
|Kit Components||96 Wells Quantity/Size|
|Aluminium pouches with a Microwell Plate coated with monoclonal antibody to human IFN-γ(8﹡12)||1 plate|
|Human IFN-γ Standard lyophilized, 4000 pg/ml upon reconstitution||2 vials|
|Concentrated Biotin-Conjugate anti-human IFN-γ monoclonal antibody||2 vials|
|Streptavidin-HRP solution||2 vials|
|Standard /sample Diluent||1 bottle|
|Biotin-Conjugate antibody Diluent||1 bottle|
|Streptavidin-HRP Diluent||1 bottle|
|Wash Buffer Concentrate 20x (PBS with 1% Tween-20)||1 bottle|
|Substrate Solution||1 vial|
|Stop Solution||1 vial|
|Adhesive Films||4 pieces|
|Product data sheet||1 copy|
|Storage||Store at 2 - 8°C|
|REPEATABILITY||The coefficient of variation of both intra-assay and inter-assay were less than 10%.|
|SENSITIVITY||The minimum detectable dose was 7pg/mL.|
|ASSAY RANGE||15.6 - 1000 pg/mL|
|SPECIFICITY||This assay recognizes both natural and recombinant human IFN-γ. The factors listed below were prepared at 10ng/ml in Standard /sample Diluent and assayed for cross-reactivity and no significant cross-reactivity or interference was observed.|
Factors assayed for cross-reactivity
|Recombinant human||EGF, IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12, MCP-1, TGF-β1, TGF-β2, TGF-β3, TGF-β5, VEGF, TNF-α, TNF-β|
|Recombinant mouse||TNF-α, IFN-γ, IL-1α, IL-1β, IL-3, IL-4, IL-5, IL-6, IL-7, IL-9, IL-10, GM-CSF, LIF|
Data Analysis Assistance
We have partnered with MyAssays to offer you an easy to use and versatile tool to analyze the data you receive using our ELISA Kit. Click the link below to be directed to the data analysis tool provided by MyAssays specifically for BSKH1013.