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Updated: Aug 9, 2026

A Rapid High-throughput Method for Mapping Ribonucleoproteins (RNPs) on Human pre-mRNA
13:00

A Rapid High-throughput Method for Mapping Ribonucleoproteins (RNPs) on Human pre-mRNA

Published on: December 2, 2009

Hapten labeling of nucleic acids for immuno-polymerase chain reaction applications

Michael Adler1

  • 1Chimera Biotech GmbH, Dortmund, Germany.

Methods in Molecular Biology (Clifton, N.J.)
|June 16, 2004
PubMed

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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