Detecting multiple proteins by Western blotting using same-species primary antibodies, precomplexed serum, and hydrogen peroxide

Ritika Upadhaya, Wataru Mizunoya, Judy E. Anderson

Research output: Contribution to journalArticlepeer-review

14 Citations (Scopus)

Abstract

Western blot detection of multiple proteins is challenged by the need to use antibodies from the same species and the harsh stripping methods that can remove protein or reduce protein antigenicity. Quenching using 27% hydrogen peroxide was developed as an alternative to stripping to inhibit horseradish peroxidase used to detect secondary antibodies. To detect two epitopes with same-species primary antibodies, quenching was followed by incubation in a precomplexed mixture of primary and secondary antibodies for the second epitope plus serum from that species. Both methods will be valuable in specific detection of multiple proteins by Western blotting, and will save time, valuable samples, and reagents.

Original languageEnglish
Pages (from-to)342-344
Number of pages3
JournalAnalytical Biochemistry
Volume419
Issue number2
DOIs
Publication statusPublished - Dec 15 2011

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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