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. Standards and samples are pipetted into the wells and any IL-1β 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-1β is added to the wells and 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-1β 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), originally described as lymphocyte activating factor (LAF) for its effects on thymocytes, is a polypeptide cytokine with two molecular forms. The two distinct molecular forms of IL-1 are thought to be derived from two genes. After transcription, as 31kD precursor polypeptide is cleaved to give rise to mostly cell membrane associated IL-1α and secreted IL-1β. Both have the same molecular weight of 15kD but have different isoelectric points of 5 and 7, respectively.
Despite sequence homology of only 20%, both forms are thought to bind to the same receptor. IL-1 inhibitors that vary only in their degree of glycosylation have been described to bind to the IL-1 receptor. These inhibitors are structurally related to IL-1β and may be important in regulation of IL-1β action.
IL-1β is produced primarily by monocytes and macrophages but also by astrocytes, oligodendroglia, adrenal cortical cells, NK cells, endothelial cells, keratinocytes, megakaryocytes, platelets, neurons, neutrophils, osteoblasts, Schwann cells, trophoblasts, T cells, and fibroblasts. IL-1 has multiple immunological functions including enhancement of IL-2 production by T cells and activation of B-cells (BAF) and thymocytes. A true pleiotrope, IL-1 may have tumoricidal activity via its release of IL-2 and Interferon gamma and indirectly antiviral by stimulating to release interferon beta.
Low levels of IL-1β have been reported in normal serum. It is thought that IL-1 genes are induced to respond to tissue damage or infection. Elevated levels have been reported in a number of infectious disease conditions and in noninfectious inflammatory conditions such as Crohn’s disease. In addition to elevated serum levels, IL-1 has been found in synovial fluids of patients with rheumatoid arthritis and in cerebrospinal fluid after neurological inflammation or insult. At the other end of the spectrum, low levels of IL-1 have been found in malnutrition and advanced neoplasia suggesting perhaps a complex immunological and physiological regulatory role for this cytokine.
|SYNONYMS||Interleukin-1 beta; IL1B; L1F2|
|Kit Components||96 Wells Quantity/Size|
|Aluminium pouches with a Microwell Plate coated with monoclonal antibody to human IL-1β(8﹡12)||1 plate|
|Human IL-1β Standard lyophilized, 500 pg/ml upon reconstitution||2 vials|
|Concentrated Biotin-Conjugate anti-human IL-1βmonoclonal 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 is less than 10%.|
|SENSITIVITY||The minimum detectable dose is 4pg/mL.|
|ASSAY RANGE||7.81- 500 pg/mL|
|SPECIFICITY||This assay recognizes both natural and recombinant human IL-1β. The factors listed below were prepared at 50ng/ml in Standard /samples. 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-1ra, IL-1 sRI, IL-1 sRII|
Data Analysis Assistance
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