[Histonet] Buffer for ISH

[Susan Bachus]

It only takes a few minutes to make some hybridization buffer up IF you make 
some 50% dextran sulfate in advance (which does take a LONG time to get into 
solution!) and keep that in the refrigerator.   I've continued to use a 
large batch of dextran sulfate for years.   If you warm the dextran sulfate 
up for an hour or so in advance (I put it in my hybridization incubator) it 
really doesn't take long to add the other ingredients for a fresh batch of 
hybridization buffer for each experiment.   The hybridization buffer can be 
kept for a while (months) in the freezer but ingredients will slowly begin 
to precipitate out.   We've consistently had good results by coasting on the 
same batch of dextran sulfate as long as it lasts but making the buffer 
fresh for each assay.    Susan
----- Original Message ----- 
From: "Liz Chlipala" <liz <@t> premierlab.com>
To: "Richard Cartun" <Rcartun <@t> harthosp.org>; "Histonet" 
<histonet <@t> lists.utsouthwestern.edu>
Sent: Thursday, February 21, 2008 12:27 PM
Subject: RE: [Histonet] Buffer for ISH


Richard

I was looking for one too, but I could not locate one.  We prepared our
own.  We buy alot of our probes from Genedetect and they were telling me
that in the future they may provide an hybidization buffer.  I use
theirs, it takes about 2 days to make because of the high concentration
of dextran sulfate.  Teri Johnson sent me one that looked a bit simpler.
I have a SOP for the one I use if you are interested.

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
P.O. Box 18592
Boulder, CO 80308
phone (303) 735-5001
fax (303) 735-3540
liz <@t> premierlab.com
www.premierlab.com

Ship to Address:

Premier Laboratory, LLC
University of Colorado at Boulder
MCDB, Room A3B40
Boulder, CO 80309

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Richard
Cartun
Sent: Thursday, February 21, 2008 10:00 AM
To: Histonet
Subject: [Histonet] Buffer for ISH

I am posting this question for a friend that works in veterinary
medicine:

"is there a commercially available hybridization buffer for ISH that can
be purchased separately (or as part of a kit)?"

Thanks,

Richard

Richard W. Cartun, Ph.D.
Director, Immunopathology & Histology
Assistant Director, Anatomic Pathology
Hartford Hospital
80 Seymour Street
Hartford, CT  06102
(860) 545-1596
(860) 545-0174 Fax

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