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

[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 
sections.

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 <gcallis <@t> montana.edu> 01/10/05 05:00PM >>>
>Liz,
>
>As discussed many times on Histonet, CD4 will NEVER work on FFPE!  FYI,
>CD8
>also will not work.
>
>If you have to run all these antibodies on the same sample, you will
>have
>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
>at
>RT next day.  Do NOT air dry again, go directly from fixative to buffer
>and
>then stain.  We actually use biotinylated primaries more than purified,
>
>then come back with Strepavidin-HRP, then DAKO AEC+ or Chris van der
>Loos
>AEC - his formulation is excellent inhouse chromogen.  Using
>biotinylated
>primaries shortens a protocol!
>
>We buy all antibodies, monoclonals from BD PharminGen.  SEROTEC is
>another
>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
>me
> >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
> >liz <@t> premierlab.com
> >www.premierlab.com
> >
> >Ship to Address:
> >Premier Laboratory
> >University of Colorado
> >MCDB, Room A3B40
> >Boulder, Colorado 80309
> >
> >_______________________________________________
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>
>Gayle Callis
>MT,HT,HTL(ASCP)
>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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