Principle of the Assay
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for IL-1β has been pre-coated onto a microplate. Standard, control, or sample and the working solution of Biotin-Conjugate are pipetted into the wells. Following incubation and wash steps, any IL-1β present is bound by the immobilized antibody and the detection antibody specific for IL-1β is binds to the combination of capture antibody- IL-1β 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-1β 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-18 standard dilutions and IL-1β sample concentration determined.
For Use with serum, plasma and cell culture supernatants. For Research Use Only. Not for use in diagnostic procedures.
Interleukin 1 (IL-1) is a name that designates two proteins, IL-1αand IL-1β, which are the products of distinct genes, but which recognize the same cell surface receptors. With the exception of skin keratinocytes, some epithelial cells, and certain cells of the central nervous system, IL-1 is not produced by the cells of healthy individuals. However, in response to stimuli such as those produced by inflammatory agents, infections, or microbial endotoxins, a dramatic increase in the production of IL-1 by macrophages and various other cell types is seen.
IL-1α and IL-1β are structurally related polypeptides that show approximately 25% homology at the amino acid level. Both are synthesized as 31 kDa precursors that are subsequently cleaved into proteins with molecular weights of approximately 17.5 kDa.
Intracellular IL-1βconsists exclusively of the 31 kDa precursor form. Extracellular IL-1βconsists of a mixture of both unprocessed and mature IL-1β. These results indicate that processing takes place subsequently to secretion and is not tightly coupled to secretion. The specific protease apparently responsible for the processing of IL-1β, designated interleukin-1β-converting enzyme (ICE), has been described.
IL-1 possesses a wide variety of biological activities. IL-1 also plays an important role in immune functions, having effects on macrophages / onocytes, Tlymphocytes, B lymphocytes, NK cells, and LAK cells. It acts on macrophages/ monocytes, inducing its own synthesis as well as the production of TNF and IL-6. It activates T cells, resulting in IL-2 production and expression of IL-2 receptors. IL-1 also induces the production of GM-CSF and IL-4 from activated T cells. It induces B cell proliferation and maturation and increased immunoglobulin synthesis.
|Kit Components||96 Wells Quantity/Size|
|Aluminium pouches with a Microwell Plate coated with antibody to mouse IL-18 (812)||1 plate|
|Mouse IL-1β standard lyophilized, 2000 pg/ml upon reconstitution||2 vials|
|Concentrated Biotin-Conjugate anti-mouse IL-1β 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||31.25-2000 pg/mL|
|SPECIFICITY||This assay recognizes both natural and recombinant mouse IL-1β. 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||IL-1α, IL-1ra, IL-1 RI, IL-1 RII|
Data Analysis Assistance
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