[Histonet] CD4, cd8

[Kristen Broomall]

I was just commenting on this statement. "As discussed many times on
Histonet, CD4 will NEVER work on FFPE!  FYI, CD8 
also will not work."

-----Original Message-----
From: Baldridge, Lee Ann [mailto:lhadley <@t> iupui.edu]
Sent: Tuesday, January 11, 2005 1:04 PM
To: Joanne Mauger; Histonet <@t> lists.utsouthwestern.edu;
gcallis <@t> montana.edu; kbroomal <@t> NEMOURS.ORG; liz <@t> premierlab.com
Subject: RE: [Histonet] CD4, cd8


Hi Everyone,
If you check the original post you'll find we're talking about mouse
tissue.  We too have great success with cd4 & cd8 but in ffpe Human
tissue not mouse.
Good Luck,
Lee Ann Baldridge
IUSM
Indpls,IN.

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Joanne
Mauger
Sent: Tuesday, January 11, 2005 12:22 PM
To: Histonet <@t> lists.utsouthwestern.edu; gcallis <@t> montana.edu;
kbroomal <@t> NEMOURS.ORG; liz <@t> premierlab.com
Subject: RE: [Histonet] CD4, cd8


To All,

We have used CD4 and CD8 from Novocastra with great success on FFPE.
Also CD4 from Zymed works on FFPE- all need EDTA retreival.
Jo



>>> "Kristen Broomall" <kbroomal <@t> NEMOURS.ORG> 01/11/05 10:18 AM >>>
Gayle,

We've been using CD4 & CD8 on FFPE tissues with not much problem.

They are from Neomarkers (www.labvision.com).

CD4 Ab-8
http://www.labvision.com/ab.cfm?first=AntiBody&second=1528 

CD8 Ab-1
http://www.labvision.com/ab.cfm?first=AntiBody&second=457 

Kristen Broomall, HT (ASCP)

-----Original Message-----
From: Gayle Callis [mailto:gcallis <@t> montana.edu] 
Sent: Monday, January 10, 2005 5:01 PM
To: Elizabeth Chlipala; Histonet <@t> lists.utsouthwestern.edu 
Subject: Re: [Histonet] CD21, B220, CD4


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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