Principle of the Assay
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for IL-2 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any IL-2 present is bound by the immobilized antibody. Following incubation unbound samples are removed during a wash step, and then a detection antibody specific for IL-2 is added to the wells and binds to the combination of capture antibody- IL-2 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 IL-2 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 IL-2 standard dilutions and IL-2 sample concentration determined.
For Use with serum, plasma and cell culture supernatants. For Research Use Only. Not for use in diagnostic procedures.
Interleukin 2 (IL-2) is a pleiotropic cytokine produced primarily by mitogen- or antigen-activated T lymphocytes. Human IL-2 (also known as T-cell growth factor) is produced by T-cells in response to antigenic or mitogenic stimulation. IL-2 is a potent lymphoid cell growth factor which exerts its biological activity primarily on T cells promoting proliferation and maturation.
IL-2 has been found to stimulate growth and differentiation of B cells, NK cells, LAK cells, monocytes, and oligodendocytes. IL-2 is involved in the treatment of cancers such as melanoma and renal cell cancer. It plays a key role in promoting the clonal expansion of antigen-specific T cells. In addition, IL-2 has also been shown to mediate multiple immune responses on a variety of cell types.
The sequence of human IL-2 cDNA predicts a 153 amino acid (aa) residue precursor glycoprotein containing a 20 aa residue signal peptide that is cleaved to form the mature protein. At the amino acid sequence level, mouse IL-2 is approximately 60% identical to human IL-2. Whereas human IL-2 is active on mouse cells, mouse IL-2 is species-specific and is inactive on human cells. The gene for IL-2 has been mapped to human chromosome 4q.
The biological effects of IL-2 are mediated by specific cell surface receptor complexes. The functional high-affinity receptor for IL-2 is composed of three distinct polypeptide chains.
IL-2 stimulates the proliferation of thymocytes; stimulates the proliferation and differentiation of activated B cells; promotes the growth, differentiation and cytocidal activity of monocytes; induces the growth of natural killer cells and stimulates cytokine production by these cells as well as the cytolytic activity of these cells; enhances the production of lymphocyte-activated killer (LAK) cells; and induces the proliferation and differentiation of oligodendrocytes.
|SYNONYMS||Interleukin 2;IL-2;T Cell Growth Factor|
|Kit Components||96 Wells Quantity/Size|
|Aluminium pouches with a Microwell Plate coated with monoclonal antibody to human IL-2 (8﹡12)||1 plate|
|Human IL-2 Standard lyophilized, 2000 pg/ml upon reconstitution||2 vials|
|Concentrated Biotin-Conjugate anti-human IL-2 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|
|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 4pg/mL.|
|ASSAY RANGE||7.81- 500 pg/mL|
|SPECIFICITY||This assay recognizes both natural and recombinant human IL-2. The factors listed below were prepared at 100ng/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||IL-2 sRα, IL-2 Rα, IL-2 Rγ, IL-3, IL-4, IL-6, IL-7, IL-8, LIF, TGF-β1, TGF-β2, TNF-α, TNF-β|
|Recombinant mouse||IL-2, IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7|
|Other proteins||bovine FGF acidic, bovine FGF basic, human PDGF, porcine PDGF, human TGF-β1, porcine TGF-β1|
Data Analysis Assistance
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