Re: BrdU: Ethanol fixation and compatibility with fluorophores

From: Anupama Ahuja <anupama.ahuja@yale.edu>
Date: Thu May 15 2008 - 08:33:07 EDT

That is a great idea, in case I can't get surface markers to work together with BrdU protocol.

Anupama Ahuja, Ph. D.
Associate Research Scientist
Dept. of Laboratory Medicine
Yale University School of Medicine
1 Gilbert  St.
TAC S541
New Haven, CT 06510

Ph: (203)-785-6557
Fax: (203)-785-5415

Gerstein, Rachel wrote:

the way around this is to first sort your cells as live cells and then do the BrDU procedure.  just be sure to optimize your BrDU for very small #s of cells first.

=======================================================
Rachel M. Gerstein, Ph.D.
Associate Professor
Department of Molecular Genetics and Microbiology
Graduate Program in Immunology/Virology
University of Massachusetts Medical School
55 Lake Avenue North
Worcester, MA 01655-0002
(508) 856-1044
(508) 856-5920 (FAX)



-----Original Message-----
From: Anupama Ahuja [mailto:anupama.ahuja@yale.edu]
Sent: Tue 5/13/2008 2:10 PM
To: cyto-inbox Subject: BrdU: Ethanol fixation and compatibility with fluorophores

Hello

I have been using the BrdU staining protocol using Ethanol, followed by
PFA/tween-20 for a while now. I know that APC and APC-cy7 Abs don't do
well with this procedure. I was wondering if anyone knows specifically
how this protocol affects PerCP, PEcy5.5, PerCP-cy5.5, PE-cy5 and/or
PE-texasred Abs? I am practically running out on colors even for our
LSR2 and have to use one of the above-mentioned colors!

I really appreciate the help....

Anu

--
Anupama Ahuja, Ph. D.
Associate Research Scientist
Dept. of Laboratory Medicine
Yale University School of Medicine
1 Gilbert  St.
TAC S541
New Haven, CT 06510

Ph: (203)-785-6557
Fax: (203)-785-5415

Received on Thu May 15 16:38:00 2008