For the quantitative determination of human Tumor Necrosis Factor-b (TNF-b) concentrations in serum, plasma, cell culture supernatant, and other biological fluids.
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for TNF-b has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any TNF-b present is bound by the immobilized antibody. After washing away any unbound substances, an enzyme-linked polyclonal antibody-enzyme specific for TNF-b 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 TNF-b 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 TNF-b 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 TNF-b concentration (pg/ml). The concentration of TNF-b in the samples is then determined by comparing the O.D. of the samples to the standard curve. This immunoassay is calibrated against natural human TNF-b (NIBSC/WHO First International Standard 87/640).
Kit Components:T9160-40C: TNFb Microtiter Plate, 1x96wells (Pre-coated with a monoclonal anti-human TNFb)T9160-40D: Pab (Biotin) Conjugate, 1x11ml (Anti-human TNFb labeled with Biotin)T9160-40E: Avidin (HRP) Conjugate, 1x14mlT9160-40F: TNFb Standard 0.8ng/vial, 2x1vialT9160-40G: Serum Assay Buffer, 1x6mlT9160-40H: Calibrator Diluent I, 1x22mlT9160-40J: Calibrator Diluent II, 1x22mlT9160-40K: Wash Buffer, 20X, 1x60mlT9160-40L: Substrate A, 1x10mlT9160-40M: Substrate B, 1x10mlT9160-4N: Stop Solution, 1x14ml
Storage and Stability:Store components at 4°C. Stable for 6 months. For maximum recovery of product, centrifuge the original vial prior to removing the cap.
