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Principle |
The qualitative detection of Rat PV antigen is based on the Double Antibody Sandwich ELISA (Enzyme-Linked Immunosorbent Assay) technique. The microplate wells are pre-coated with an antibody specific to Rat PV. The corresponding antigen in the sample or standards binds the immobilized antibody after incubation. After washing the microplate wells to remove all unbound sample material, horseradish peroxidase (HRP) labeled antibody conjugate is added, which binds to the antigen, forming antibody-antigen enzyme-labeled antibody complex upon incubation. In a second wash step, all unbound material is removed. The immune complex formed by the bound conjugate is visualized by adding Tetramethylbenzidine (TMB) substrate which gives a blue color reaction in wells containing the complex. A stop solution containing acid terminates the reaction, producing a yellow end-point coloration proportional to the amount of target antigen present in the sample. Absorbance is measured at 450 nm using an ELISA microwell plate reader. Analysis is completed by comparing the OD values of samples to a cut-off value |
