Principle of the Assay
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for IL-15 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any IL-15 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-15 is added to the wells and binds to the combination of capture antibody- IL-15 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-15 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-15 standard dilutions and IL-15 sample concentration determined.
For Use with serum, plasma and cell culture supernatants. For Research Use Only. Not for use in diagnostic procedures.
Interleukin 15 (IL-15) is a 14 kDa cytokine that is structurally and functionally related to IL- 2. Mature human IL-15 shares 70% amino acid sequence identity with mouse and rat IL-15. Alternate splicing generates isoforms of IL-15 with either a long or short signal peptide (LSP or SSP), and the SSP isoform is retained intracellularly. IL-15 associates with IL-15 Rα in the endoplasmic reticulum, and this complex is expressed on the cell surface. The dominant mechanism of IL-15 action is known as transpresentation in which IL-15 and IL-15 Rα are coordinately expressed on the surface of one cell and interact with complexes of IL-2 Rβ/γc on adjacent cells. This enables cells to respond to IL-15 even if they do not express IL-15 Rα. Consistent with its shared use of IL-2 receptor subunits, IL-15 induces IL-2-like effects in lymphocyte development and homeostasis. It is particularly important for the maintenance and activation of NK cells and CD8+ memory T cells. IL-15 also exerts pleiotropic effects on other hematopoietic cells and nonimmune cells. Ligation of membrane associated IL-15/IL-15Rα complexes induces reverse signaling that promotes cellular adhesion, tyrosine phosphorylation of intracellular proteins, and cytokine secretion by the IL-15/IL-15Rα expressing cells.
|Kit Components||96 Wells Quantity/Size|
|Aluminium pouches with a Microwell Plate coated with monoclonal antibody to human IL-15 (8﹡12)||1 plate|
|Human IL-15 Standard lyophilized, 1000 pg/ml upon reconstitution||2 vials|
|Concentrated Biotin-Conjugate anti-human IL-15 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 2pg/mL.|
|ASSAY RANGE||7.81 - 500 pg/mL|
|SPECIFICITY||This assay recognizes both natural and recombinant human IL-15. The factors listed below were prepared at 50ng/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-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-12, IL-13, MIP-1α, MIP-1β, IFN-γ, TNF-α, TNF-β|
|Recombinant mouse||IL-1α, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-7, IL-9, IL-10, IL-13, MIP-1α, MIP-1β, SCF, TNF-α|
|Other proteins||bovine FGF acidic, bovine FGF basic, human PDGF, porcine PDGF, human TGF-β1, porcine TGF-β1, bovine FGF acidic, bovine FGF basic|
Data Analysis Assistance
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