Methanol peroxidase blocking, other protocols now in use

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From:Gayle Callis <uvsgc@msu.oscs.montana.edu>
To:antje.marcantonio@pharma.Novartis.com, histonet@pathology.swmed.edu
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Happy to hear the F4/80 worked for you. 

There is always a chance methanol peroxidase blocking will not work well
with CD markers, murine, and it is nice to know it worked with this
antibody.  If you don't get staining with a CD marker, then you might
suspect the methanol.  These caveats are passed on by reputable companies
(PharMingen and Biogenex) and may need to be heeded at some time or another.  

I have now gone to the glucose oxidase/glucose/sodium azide method (called
GLUOX in my lab!) with different drying/fixation protocol for peroxidase
blocking on murine frozen sections.  GLUOX gives far cleaner and total
quenching results, plus morphology has greatly improved. It takes more
time, but the results are worth the effort. I can thank Barb Wright for
this info, she does the GLUOX routinely on FROZEN sections.  Had done GLUOX
in the past but set aside for time considerations - WRONG CHOICE!



   

 

At 10:02 AM 8/24/00 +0100, you wrote:
>Thanks to everybody who contributed to our new protocol for F4/80 which
>works now very nicely both on paraffin and frozen sections. Special thanks
>to Gayle Callis and Cynthia Favara.
>Gayle, I almost took over your protocol, but I can tell you that I had good
>results even with methanol peroxidase blocking.
>
>Have a very nice day !
>
>Antje Marcantonio
>Pharma Novartis AG
>Basle, Switzerland
>
>
>
>
Gayle Callis
Veterinary Molecular Biology
Montana State University
Bozeman MT 59717-3610
406 994-4705
406 994-4303



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