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  • Title: Hapten labeling of nucleic acids for immuno-polymerase chain reaction applications.
    Author: Adler M.
    Journal: Methods Mol Biol; 2004; 283():163-80. PubMed ID: 15197309.
    Abstract:
    A method for the ultrasensitive protein detection in the range of 0.01 to 10,000 amol of the model antibody anti-mouse-IgG from rabbit is described, using a combination of Immuno-polymerase chain reaction (PCR) and PCR-enzyme-linked immunosorbent assay (PCR-ELISA). The antibody was first immobilized on antigen-coated microplates; in a second step, a commercially available DNA-labeled species-specific antibody was added; and finally the deoxyribonucleic marker was amplified in a PCR step, including twofold labeling with biotinylated primer and a hapten-coupled nucleotide during PCR. Subsequently, the labeled PCR product was immobilized on streptavidin-coated microplates and detected with an antibody-enzyme conjugate. The protocol could easily be adapted to the detection of other antibodies or antigens by exchanging the antigen-specific antibody. Several modifications of the method as well as optimization steps, potential error sources, and countermeasures are discussed.
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