Re: [Histonet] Antibodies for frozen mouse sections

From:Gayle Callis

Once again, you can buy these antibodies from BD Pharmingen or SEROTEC - 
however you need to determine the working concentration of your primaries 
using a dilution panel starting at 10 ug/ml - no different than for CD4, 
CD8, etc.  and do this in your own laboratory working conditions/buffers, 

We double immunofluorescence with MadCAM and PNAd so we can see 
colocalization of these markers in NALT, and lymph nodes located in 
neck/head region.  Mesenteric lymph nodes is the positive control for these 
two antibodies.

We do NOT use FFPE for any of these antibodies - frozen sections fixed with 
an acetone/alcohol fixation is wonderful for MadCAM, PNAd and probably the 
other markers as well.

At 10:00 AM 11/2/2005, you wrote:
>Hi, I am staining some frozen mouse NALT and cervical lymph node
>sections with a number of antibodies, CD4, CD8, B220, CD11c (all from
>Pharmingen) and F4/80 (Serotec) and the staining is working fine (thanks
>to Gayle Callis!!). However I want to start staining for some more cell
>markers and I was wondering if anybody could give me details of primary
>antibodies (dilutions etc) and where you get them from, that are good
>for staining natural killer (NK) cells, ICAM-1 (CD54), CD5, MadCAM-1,
>VCAM-1, PNAd and a neutrophil marker, i.e. Ly-6G.
>Thanks very much in advance.
>Lindsay Hall
>Wellcome Trust Sanger Institute
>Wellcome Trust Genome Campus
>CB10 1SA
>Histonet mailing list

Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367
406 994-4303 (FAX)

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