Principle of the Assay
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for IL-5 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any IL-5 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-5 is added to the wells and binds to the combination of capture antibody-IL-5 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-5 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-5 standard dilutions and IL-5 sample concentration determined.
For Use with serum, plasma and cell culture supernatants. For Research Use Only. Not for use in diagnostic procedures.
Mouse interleukin 5 (mIL-5), also known as T-cell replacing factor, B-cell growth factor II, eosinophil differentiation factor and eosinophil colony stimulating factor, is a pleiotropic cytokine produced primarily by T cells. It supports the proliferation and differentiation of mouse, but not human, B cells, and enhances IgM, IgG1, IgA, and IgE secretion. IL-5 chemo attracts and enhances the survival and the effector functions of mature eosinophils and synergizes with various colony stimulating factors to increase eosinophil progenitor production and eosinophil expansion. Because of its diverse effects on eosinophils, IL-5 is strongly implicated in the pathogenesis of asthma and other hypereosinophilic inflammatory conditions. Mouse IL-5 cDNA encodes a 133 amino acid (aa) residue precursor protein containing a hydrophobic signal peptide that is cleaved to yield a 113 aa residue mature protein. Native mouse IL-5 is a disulfide-linked homodimeric 40-45 kDa glycoprotein with N- and O-linked carbohydrate chains. Mature human IL-5 is approximately 70% identical at the amino acid level to mouse IL-5. Whereas mouse and human IL-5 are equally active on human cell lines, human IL-5 is much less active than mouse IL-5 in mouse cell assays. IL-5 is a main regulator of eosinopoiesis, eosinophil maturation and activation. The elevated production of this cytokine is reported to be related to asthma or hypereosinophilic syndromes.
|Kit Components||96 Wells Quantity/Size|
|Aluminium pouches with a Microwell Plate coated with monoclonal antibody to mouse IL-5(8﹡12)||1 plate|
|Mouse IL-5 Standard lyophilized, 1000 pg/ml upon reconstitution||2 vials|
|Concentrated Biotin-Conjugate anti-mouse IL-5 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 mouse IL-5. 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 mouse||G-CSF, GM-CSF, IL-1α, IL-1β, IL-2 , IL-3, IL-4, IL-6, IL-7, IL-9, IL-10, IL-12, IL-17, LIF, Leptin, TNF-α, VEGF|
|Recombinant human||IL-5, IL-5sR|
Data Analysis Assistance
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