Rat IL-1β ELISA Kit
Interleukin-1 (IL-1), originally described in 1972 as lymphocyte activating factor (LAF) for its effects on thymocytes, is a polypeptide cytokine with two molecular forms. Both forms appear to mediate identical ranges of biological activity which include synthesis of the acute phase proteins by hepatocytes, chemotaxis of polymorphonucleocytes, and release of polymorphonucleocytes from blood and bone marrow. These effects coined the acronym leukocyte endogenous mediator (LEM). Early researchers also called IL-1β endogenous pyrogen, and it has been shown to induce fever and is thought to contribute to wasting of muscles (PIF, proteolysis inducing factor). Other activities associated with IL-1 are the induction of Prostaglandin E2 by synovial cells and release of collagenase with resulting destruction of cartilage and bone resorption (catabolin, osteoclast activation factor). In addition, IL-1, has multiple immunological functions including enhancement of IL-2 production by T cells and activation of B-cells (BAF) and thymocyte. A true pleiotrope, IL-1 may have tumoricidal activity via its release of IL-2 and interferon gamma and be indirectly antiviral by stimulating fibroblasts to release interferon beta. In its role as mediator of sepsis, IL-1 has most recently been described as enhancing the growth of virulent E.coli.
An anti-rat IL-1β polyclonal coating antibody is adsorbed onto microwells.
Rat IL-1β present in the sample or standard binds to antibodies adsorbed to the microwells; a biotin-conjugated polyclonal anti-rat IL-1β antibody is added and binds to rat IFNγ captured by the first antibody.
Following incubation unbound biotin conjugated anti-rat IL-1β is removed during a wash step. Streptavidin-HRP is added and binds to the biotin conjugated anti-rat IL-1β.
Following incubation unbound Streptavidin-HRP is removed during a wash step, and substrate solution reactive with HRP is added to the wells.
A coloured product is formed in proportion to the amount of rat IL-1β present in the sample.
The reaction is terminated by addition of acid and absorbance is measured at 450 nm.
A standard curve is prepared from seven rat IL-1β standard dilutions and rat IL-1β sample concentration determined.