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Pre-hybridisation: an efficient way of suppressing endogenous biotin-binding activity inherent to biotin-streptavidin detection system

  • Raju Ahmed
    ,
  • Emma Spikings
    ,
  • Shaobo Zhou
    ,
  • Andrew Thompsett
    ,
  • Tiantian Zhang
  • University of East London
    ,
  • Bournemouth University
Research Output: Contribution to journal Article Peer-review

Open access

Abstract

Endogenous biotin or biotinylated protein binding activity is a major drawback to biotin-avidin/streptavidin detection system. The avidin/streptavidin conjugate used to detect the complex of the biotinylated secondary antibody and the primary antibody binds to endogenous biotin or biotinylated proteins leading to non-specific signals. In Western blot, the endogenous biotin or biotinylated protein binding activity is usually manifested in the form of ~72kDa, ~75kDa and ~150kDa protein bands, which often mask the signals of interest. To overcome this problem, a method based on prior hybridisation of the biotinylated secondary antibody and the streptavidin conjugate was developed. The method was tested alongside the conventional biotin-streptavidin method on proteins extracted from zebrafish (Danio rerio) embryos. Results showed that the newly developed method efficiently suppresses the endogenous biotin or biotinylated protein binding activity inherent to the biotin-streptavidin detection system.

Publication Information

Output type

Research Output: Contribution to journal Article Peer-review

Original language

English

Pages from-to (Number of pages)

Pages 143-147

Journal (Volume, Issue Number)

Journal of Immunological Methods (Volume 406)

Publication milestones

  • Accepted/In press - 11/03/2014
  • Published - 18/03/2014

Publication status

Published - 18/03/2014

ISSN

0022-1759

External Publication IDs

  • handle.net: 10547/560960
  • Scopus: 84900849843

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