Re: need help F4/80


we use the F4/80 antibody from Serotec for both frozen and formalin fixed paraffin sections.
For this antibody the fixation mustn't be longer than 2-3 days in formalin. This is true for an optimal staining for many antibodies, but with the F4/80 it is really obligatory.
I never tried any boosting here because there is always a little bit of background, which we reduced to a minimum by adding mouse serum to the blocking solution.
Pretreatment: 0.2% trypsin for 10 min
We use the primary 1:100 diluted in the blocking solution (10% normal rabbit serum plus 2.5 % mouse serum) , incubation overnight (for frozens 60 min, rt).
As secondary: rabbit anti-rat, mouse adsorbed , Vector
I have to admit that staining of frozen sections is less problematic but we are happy with our staining on FFPE , too.
Good luck !

Antje Marcantonio
Novartis Pharma AG
BU Transplantation Research

Robert Schoonhoven <>

09.01.2003 13:20

        cc:        "''" <>
        Subject:        need help F4/80

Mouse IHC folks:

Has anyone had any luck with the rat anti mouse F4/80 (Accurate Scientific)?
This one just doesn't want to work for me on formalin fixed paraffin sections.

In brief I've tried the following:

Pretreatment with trypsin, HEIR citrate, and proteinase

primary dilution's of 1:50, 100 & 500

If anyone has had any good results using this antibody please contact me.


Robert Schoonhoven
Laboratory of Molecular Carcinogenesis
University of North Carolina


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