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

Regards

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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