[Histonet] Flippin on the Bond

Tony Henwood (SCHN) tony.henwood at health.nsw.gov.au
Mon Mar 16 18:18:36 CDT 2020

Hi all,

I was reading an article entitled “Evaluation of Tissue Adhesion and Staining Performance of BOND Adhesive Microscope Slides: A Comparative Study on the BOND-III and BOND-MAX Platforms” (https://pdfs.semanticscholar.org/f5f1/ad0741bc5445d0e1079a0d07a32c8a3a26c1.pdf) and they make mention of the “Flippin Protocol”, an amended protocol widely used by BOND customers.
It is used for those antibodies that are adversely affected by the endogenous peroxidase block that is used in the Bond kits (and possibly with other non-Bond kits as well).
In this protocol, the peroxidase block is applied after the localisation antibody is applied. Examples given by Leica are PAX5 and AMACR.

The paper refers to several other antibodies as well (CD3, CD34, SMA and TTF1) that uses the “Flippin protocol”. Now I have not had issues with the first three antibodies (they work quite well with the standard protocol, ie peroxidase block before the localisation antibody) and have no experience with TTF1 (we are a pediatric laboratory).

Has anyone experienced this issue with antibodies and use the Flippin protocol?

What antibodies are an issue?

Thanks for your input


Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) | Principal Scientist; Adjunct Fellow, School of Medicine, University of Western Sydney; Visiting Lecturer, School of Life Sciences, Faculty of Science, University of Technology Sydney | Histopathology
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