Description
Intended Uses: The IL-8 ELISA Set is intended for use in a 'do it yourself' solid phase sandwich ELISA for the in vitro qualitative and quantitative determination of IL-8 in supernatants, buffered solutions, serum, plasma samples and other body fluids. This assay will recognise both natural and recombinant human IL-8.
Principle of the Assay: A capture Antibody highly specific for IL-8 is coated to the wells a microtitre strip plate. Binding of IL-8 samples and known standards to the capture antibodies and subsequent binding of the biotinylated anti-IL- 8 secondary antibody to the analyte is completed during the same incubation period. Any excess unbound analyte and secondary antibody is removed. The HRP conjugate solution is then added to every well including the zero wells, following incubation excess conjugate is removed by careful washing. A chromogen substrate is added to the wells resulting in the progressive development of a blue coloured complex with the conjugate. The colour development is then stopped by the addition of acid turning the resultant final product yellow. The intensity of the produced coloured complex is directly proportional to the concentration of IL-8 present in the samples and standards. The absorbance of the colour complex is then measured and the generated OD values for each standard are plotted against expected concentration forming a standard curve. This standard curve can then be used to accurately determine the concentration of IL-8 in any sample tested