IL-1 Beta Human ELISA Assay
The IL-1 Beta Human ELISA Assay is For Research Use Only
Size: 1×96 wells
Sensitivity: 2 pg/mL
Dynamic Range: 7.8 – 250 pg/ml
Incubation Time: 3.5 hours
Sample Type: Serum, Plasma, Cell Culture
Sample Size: 100 µl
Alternative Names: Interleukin-1 Beta, IL-1B, IL-1β
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-1a and secreted IL-1b. 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-1b action.
IL-1 Beta 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-1 Beta have been reported in normal serum . It is thought that IL-1 genes are induced to respond to tissue damage or in 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.
Related Products to IL-1 Beta Human ELISA Assay
Human IL-2 ELISA Assay Kit
Human IL-4 ELISA Assay Kit
Human IL-5 ELISA Assay Kit
The Human Interleukin 1 Beta (IL-1β) ELISA Assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for IL-1b has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any IL-1b 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-1βis added to the wells and binds to the combination of capture antibody-IL-1βin 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 colored 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.
- Prepare all reagents and working standards as directed in the previous sections.
- Determine the number of microwell strips required to test the desired number of samples plus appropriate number of wells needed for running blanks and standards. Remove extra microwell strips from holder and store in foil bag with the desiccant provided at 2-8°C sealed tightly.
- Add 100µL of Standard, control, or sample, per well. Cover with the adhesive strip provided. Incubate for 1.5 hours at 37°C.
- Aspirate each well and wash, repeating the process three times for a total of four washes. Wash by filling each well with Wash Buffer (350 µL) using a squirt bottle, manifold dispenser or auto-washer. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.
- Add 100 µL of the working solution of Biotin-Conjugate to each well. Cover with a new adhesive strip and incubate 1 hour at 37°C.
- Repeat the aspiration/wash.
- Add 100 µL of the working solution of Streptavidin-HRP to each well. Cover with a new adhesive strip and incubate for 30 minutes at 37°C Avoid placing the plate in direct light.
- Repeat the aspiration/wash.
- Add 100 µL of Substrate Solution to each well. Incubate for 10-20 minutes at 37C. Avoid placing the plate in direct light.
- Add 100 µL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
- Determine the optical density of each well immediately, using a microplate reader set to 450 nm. (optionally 630nm as the reference wave length;610-650nm is acceptable)
Typical Standard Curve
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