[Histonet] RE: F4/80

[Melissa Gonzalez]

Hi Susan, 
I am working on the same thing right now. F4/80 from Serotec used to
work fantastically, and for some reason the last few batches have been
giving me trouble (from this year). The DOL is just not anywhere what is
used to be, and this was a solid marker. I have worked around it by
changing up antigen retrievals, but it still is finicky for each
different study now. 
If you wish to use it for fluorescence, use either Molecular Probes AF
488 (green like FITC) or AF594 (red like Texas Red). The Goat anti Rat
for 30 minutes at 1:200 RT works like a charm. (for either frozen or
paraffin sections). Mount in a good antifade, and you are all set. 
I'm actually switching back and forth between CalTag's F4/80 and
Serotec's now, depending on whichever one gives me the best result.
Neither is as good as the last "working" lot of Serotec's. I was curious
if others are also are experiencing this problem...

Thanks and good luck Susan, 
Melissa


Message: 13
Date: Tue, 7 Nov 2006 14:42:31 -0000
From: <Susan.Ferrigon <@t> sanofi-aventis.com>
Subject: [Histonet] F4/80
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<90B6684A9D6DAF468F7A5DC148754E1D1E1869 <@t> ALPW31.f2.enterprise>
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Hi Histonetters
 
I have been able to stain some mouse liver for Kuppfer cells using
Serotec's F4/80 and copious amounts of help from their tech support,
however I would like to use some immunofluorescence instead of the DAB.
At present I am using a Goat anti Rat IgG( Mouse adsorbed) :HRP
secondary.  Does anyone know of another secondary I could use with a
fluorescent marker.  I am using FFPE or cryostat sections.
 
Thanks
 
Susan