Interleukin-1 alpha(IL-1a) is a proinflammatory cytokine produced by a wide variety of cell types, including macrophages, osteoblasts, monocytes and hepatocytes. IL-1 alpha is synthesized as 31 kDa precursors that are subsequently cleaved into proteins with molecular weights of approximately 17 kDa. The IL-1a precursor reportedly shows full biological activity in the EL-4 assay. Circulating levels of IL-1a are normally low and only rise after stimulation by agents such as those produced byinflammation, infection or microbial endotoxins. IL-1a possesses a wide variety of biological activities and exerts its effects by binding to specific cell surface receptors.

This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for IL-1α has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any IL-1α 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-1α is added to the wells and binds to the combination of capture antibody-IL-1α 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-1α 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-1α standard dilutions and IL-1α sample concentration determined.

Figure 1:Schematic diagram of the assay

1. Aluminium pouches with a Microwell Plate coated with antibody to mouse IL-1α (812)

2. 2 vials mouse IL-1α Standard lyophilized, 500 pg/ml upon reconstitution

3. 2 vials concentrated Biotin-Conjugate anti-mouse IL-1α antibody

4. 2 vials Streptavidin-HRP solution,

5. 1 bottle Standard /sample Diluent

6. 1 bottle Biotin-Conjugate antibody Diluent

7. 1 bottle Streptavidin-HRP Diluent

8. 1 bottle Wash Buffer Concentrate 20x (PBS with 1% Tween-20)

9. 1 vial Substrate Solution

10. 1 vial Stop Solution

11. 4 pieces Adhesive Films

12. package insert

NOTE: [96 Tests]

Unopened Kit：Store at 2 -8° C. Do not use past kit expiration date. opened/ReconstitutedReagents：Please refer to the datasheets for detail information.

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