Capture Antibody: Pre-titrated, purified antibody
Detection Antibody: Pre-titrated, biotin-conjugated antibody
Standard: Recombinant protein for generating standard curve and calibrating samples
10X Coating Buffer: Buffer for plating the Capture Antibody
5X ELISA/ELISPOT Diluent: Buffer for blocking and diluting the Detection Antibody and Enzyme
Enzyme: Pre-titrated Avidin-HRP
Substrate: 1X TMB Solution
Certificate of Analysis: Lot-specific instructions for dilution of antibodies and standards
96 Well Plate: Corning Costar 9018 (included with product Cat. Nos. ending in suffixes -22, -76, -86)

The Mouse Interleukin-1beta (IL-1b) Uncoated ELISA Kit contains pre-matched antibody pairs, and reagents for performing quantitative enzyme linked immunosorbent assays (ELISA) to detect and quantify protein levels of mouse IL-1b. Wash Buffer and Stop Solution are needed to complete the ELISA reaction and are sold separately.

Principle of the method

ELISAs are designed to measure the amount of the target bound between a matched antibody pair. A target-specific antibody is coated to the bottom of the wells of a microplate, which is an overnight process. Samples, standards, or controls are then added into these wells and bind to the immobilized (capture) antibody. A sandwich is formed by the addition of the second (detector) antibody, a substrate solution is added that reacts with the enzyme-antibody-target complex to produce measurable signal. The intensity of this signal is directly proportional to the concentration of target present in the original specimen.

Interleukin-1 beta (IL-1 beta) is a proinflammatory cytokine expressed by monocytes, macrophages, and dendritic cells. IL-1 beta is synthesized in response to inflammatory stimuli as a 31 kDa inactive pro-form that accumulates in the cytosol. Cleavage of pro-IL-1 beta into the active 17 kDa protein requires the activation of inflammasomes, which are multi-protein complexes that respond to pathogens, stress conditions, and other danger signals. Inflammasome activation triggers the processing of the caspase-1 precursor into its active form, which in turn cleaves pro-IL-1 beta. IL-1 beta lacks a signal sequence peptide for classical ER/Golgi pathway and is secreted alongside caspase-1 via an alternate and incompletely understood mechanism. Although IL-1 beta is most often secreted in its active form, secretion of the uncleaved protein may be detectable under some biological conditions. IL-1 beta signals through two receptors, IL-1RI and IL-1RII, both of which are shared with IL-1 alpha. IL-1 beta activity can be moderated by IL-1 Receptor Antagonist (IL-1RA), a protein produced by many cell types that blocks receptor binding through competitive inhibition. IL-1 beta play an important role in innate host defense by triggering the production of other proinflammatory cytokines in target cells and initiating acute-phase responses to infection and injury. Elevated levels of IL-1 beta have been associated with many chronic inflammatory conditions IL-1 beta neutralizing antibodies potential therapeutic value.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.