Question was responded to about antibodies for MOUSE tissue [Histonet] CD21, B220, CD4

From:Gayle Callis

To set the record straight, Liz was asking about doing CD4, and the other 
markers on MOUSE tissue using FFPE.

The answer is - no, CD4 does not work on murine tissue FFPE, only frozen 

Somehow human tissue crept into this messaging which we know works very 
well with CD4 and CD8.  Doggone mouse is a problem however.

At 08:29 AM 1/11/2005, you wrote:
>CD4 and CD8 work for us.
>CD4 (Novacastra) antigen retreived in citrate buffer, or trilogy work
>fine. CD8(Dako) antigen retreive in citrate buffer.
>Doug Geddes BSc, MLT
>London Health Sciences Centre
>London Ontario, Canada
> >>> Gayle Callis  01/10/05 05:00PM >>>
>As discussed many times on Histonet, CD4 will NEVER work on FFPE!  FYI,
>also will not work.
>If you have to run all these antibodies on the same sample, you will
>to do frozen sections in order to accomadate the CD4.  FS are air dried
>overnight at RT over dessicant.   To improve morphology, use 75%
>acetone/25% absolute ethanol (no substitute on alcohol here) for 5 min
>RT next day.  Do NOT air dry again, go directly from fixative to buffer
>then stain.  We actually use biotinylated primaries more than purified,
>then come back with Strepavidin-HRP, then DAKO AEC+ or Chris van der
>AEC - his formulation is excellent inhouse chromogen.  Using
>primaries shortens a protocol!
>We buy all antibodies, monoclonals from BD PharminGen.  SEROTEC is
>supplier.  These are rat antimouse monoclonals.
>At 01:59 PM 1/10/2005, you wrote:
> >I need to run these antibodies on mouse tissue.  Can anyone recommend
> >a good source and if they might possibly work in FFPE tissue or do I
> >need to use frozens.
> >
> >Thanks
> >
> >Liz
> >
> >Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
> >Premier Laboratory, LLC
> >P.O. Box 18592
> >Boulder, Colorado 80308
> >Office: (303) 735-5001
> >Fax: (303) 735-3540
> >
> >
> >
> >Ship to Address:
> >Premier Laboratory
> >University of Colorado
> >MCDB, Room A3B40
> >Boulder, Colorado 80309
> >
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> >
>Gayle Callis
>Research Histopathology Supervisor
>Veterinary Molecular Biology
>Montana State University - Bozeman
>PO Box 173610
>Bozeman MT 59717-3610
>406 994-6367 (lab with voice mail)
>406 994-4303 (FAX)
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