Description
Intended Uses: The CD62E ELISA Set is intended for use in a 'do it yourself' solid phase sandwich ELISA for the in vitro qualitative and quantitative determination of soluble Endothelial Leucocyte Adhesion Molecule ELAM-1, (CD62E) in supernatants, buffered solutions, serum, plasma samples and other body fluids. This assay will recognise both natural and recombinant human CD62E.
Principle of the Assay: A capture Antibody highly specific for CD62E is coated to the wells a microtitre strip plate. Binding of CD62E in samples and known standards to the capture antibodies is completed and then any excess unbound analyte is removed. During the next incubation period the binding of the biotinylated anti-CD62E secondary antibody to the analyte occurs. Any excess unbound secondary antibody is then 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 CD62E 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 CD62E in any sample tested