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Direct Peroxidase Labeling of Hybridization Probes and Chemiluminescence Detection

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Nonradioactive Analysis of Biomolecules

Part of the book series: Springer Lab Manuals ((SLM))

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Abstract

The labeling of long nucleic acid probes is based on the work first described by Renz at the European Molecular Biology Laboratories (EMBL) (Renz and Kurz, 1984). Single stranded nucleic acid (RNA or DNA) is labeled with a positively-charged, modified, horseradish peroxidase (HRP) complex in a rapid, reliable and simple reaction process to produce a stable probe. In conjunction with enhanced chemiluminescence (ECL), a light-based detection system, the HRP-labeled probes can be used to detect single copy genes in as little as 1.ig of a restriction enzyme digest of human DNA blotted onto either nylon or nitrocellulose membranes (Stone and Durrant, 1991). The light output of the ECL reaction is captured on X-ray film; for most high sensitivity applications the exposure time required is less than 60 min, although up to 4-h exposures are possible. For high target applications the associated rapid hybridization and rapid detection procedures enable the whole process to be completed in 1 working day.

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Author information

Authors and Affiliations

  1. Amersham Pharmacia Biotech, Amersham Laboratories, White Lion Road, Amersham, Buckinghamshire, HP7 9LL, UK

    Ian Durrant

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  1. Ian Durrant
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Editor information

Editors and Affiliations

  1. Roche Molecular Systems Werk Tutzing, Roche Diagnostics GmbH, Bahnhofstrasse 9-15, 82327, Tutzing, Germany

    Christoph Kessler

  2. Gene Center/Institute of Biochemistry, Ludwig-Maximilians-Universität München, Feodor-Lynen-Str. 25, 81377, München, Germany

    Christoph Kessler

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© 2000 Springer-Verlag Berlin Heidelberg

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Durrant, I. (2000). Direct Peroxidase Labeling of Hybridization Probes and Chemiluminescence Detection. In: Kessler, C. (eds) Nonradioactive Analysis of Biomolecules. Springer Lab Manuals. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-57206-7_12

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  • DOI: https://doi.org/10.1007/978-3-642-57206-7_12

  • Publisher Name: Springer, Berlin, Heidelberg

  • Print ISBN: 978-3-540-64601-3

  • Online ISBN: 978-3-642-57206-7

  • eBook Packages: Springer Book Archive

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