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