Automated sequential chromogenic IHC double staining with two HRP substrates

PLoS One. 2018 Nov 20;13(11):e0207867. doi: 10.1371/journal.pone.0207867. eCollection 2018.

Abstract

Automated IHC double staining using diaminobenzidine and HRP Magenta is illustrated utilizing a new acidic block with sulfuric acid to prevent cross-reactivity. Residual cross-reactivity in double staining is determined to arise from chromogenic-bound antibodies and amplification system during the first part of the double staining.

MeSH terms

  • Automation
  • Benzidines / metabolism
  • Diazonium Compounds / metabolism
  • Horseradish Peroxidase / metabolism*
  • Immunohistochemistry / methods*
  • Staining and Labeling / methods*

Substances

  • Benzidines
  • Diazonium Compounds
  • Fast Red B
  • benzidine
  • Horseradish Peroxidase

Grants and funding

The authors received no specific funding for this work. All authors are employed by Agilent Technologies. Agilent Technologies provided support in the form of salaries for all three authors, but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the ‘author contributions’ section.