This Human Interleukin-1a ELISA Kit is to be used for the in vitro quantitative determination of human interleukin-1a (IL-1a) concentrations in serum, plasma, cell culture supernatant, and other biological fluidsIL-1alpha is a non-secreted proinflammatory cytokine produced in a variety of cells including monocytes, tissue macrophages, keratinocytes and other epithelial cells. Both IL-1alpha and IL-1beta binds to the same receptor and has similar if not identical biological properties. These cytokines have a broad range of activities including, stimulation of thymocyte proliferation, by inducing IL-2 release, B-cell maturation and proliferation, mitogenic FGF-like activity and the ability to stimulate the release of prostaglandin and collagenase from synovial cells. However, whereas IL-1beta is a secreted cytokine, IL-1alpha is predominantly a cell-associated cytokine.This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for IL-1a has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any IL-1a present is bound by the immobilized antibody. After washing away any unbound substances, an enzyme-linked polyclonal antibody-enzyme specific for IL-1a is added to the wells. Following a wash to remove any unbound antibody-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of IL-1a bound in the initial step. The color development is stopped and the intensity of the color is measured.In order to measure the concentration of IL-1a in the samples, this kit includes two calibration diluents (Calibrator Diluent I for serum/plasma testing and Calibrator Diluent II for cell culture supernatant testing.) According to the testing system, the provided standard is diluted (2-fold) with the appropriate Calibrator Diluent and assayed at the same time as the samples. This allows the operator to produce a standard curve of Optical Density (O.D) versus IL-1a concentration (pg/ml). The concentration of IL-1a in the samples is then determined by comparing the O.D. of the samples to the standard curve.Sensitivity:Calibrator Diluent I: 5.0pg/mlCalibrator Diluent II: 4.6pg/ml Range:15.6-500pg/mlKit Components:I7663-02B1:Microtiter Plate 1x96 wellsI7663-02B2: IL-1a (Biotin) Mab 1x11mlI7663-02B3: Avidin (HRP) 1x14ml*I7663-02B4: IL-1a Standard 2x1 vialI7663-02B5: Calibrator Diluent I 1x22mlI7663-02B6: Calibrator Diluent II 1x22ml.I7663-02B7: Wash Buffer (20X) 1x60mlI7663-02B8:. Substrate A 1x10mlI7663-02B9: Substrate B 10mlI7663-02B10: Stop Solution 1x14mlStorage and Stability:Store at 4°C. Store I7663-02B4: IL-1a Standard powder at 4°C, liquid at -20°C. Stable for 6 months. For maximum recovery of product, centrifuge the original vial prior to removing the cap.