[Histonet] RE: Look at me...

[Connolly, Brett M]

I'll second that from our experience - Cell Signaling cleaved caspase-3 antibody works well on xenografts

Brett

Brett M. Connolly, Ph.D.
Imaging Research Fellow
Merck & Co., Inc.
PO Box 4, WP-44K
West Point, PA 19486
brett_connolly <@t> merck.com
T- 215-652-2501
F- 215-993-6803


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Elizabeth Chlipala
Sent: Thursday, January 05, 2012 12:36 PM
To: 'Sarah Dysart'; histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] RE: Look at me...

If you are looking to stain cleaved caspase 3 there is a better antibody from cell signaling it works in multiple species, it's a bit pricey but I have found that it works the best out of the few that I have tried.  We have used it mouse xenografts before without any issue.

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
PO Box 18592
Boulder, CO 80308-1592
(303) 682-3949 office
(303) 682-9060 fax
(303) 881-0763 cell
www.premierlab.com

Ship to address:

1567 Skyway Drive, Unit E
Longmont, CO 80504

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Sarah Dysart
Sent: Thursday, January 05, 2012 10:17 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Look at me...

I'm just full of questions today!!  This one is IHC...I have been trying to optimize a Caspase3 stain for several months now and it is still just chalked full of background gradoo.  I do all the blocking including Fc receptors...still junk.  The clone I have been using is, from abcam (ab2302).  I don't see the specific clone name listed.  I am staining human xenografts raised in mouse.  I get a whole lot of background staining making it very hard to find the positive staining.  The recommended dilution is about 1:30, but I have diluted all the way up to 1:500.  At the higher dilution no positive staining or background is observed.  Does anyone know of a good Caspase3 antibody,  preferably mouse monoclonal?  All the rabbit polyclonal antibodies are difficult to stain on these xenografts.
Thanks again =)

Sarah Goebel-Dysart, BA, HT(ASCP)
Histotechnologist
Mirna Therapeutics
2150 Woodward Street
Suite 100
Austin, Texas  78744
(512)901-0900 ext. 6912

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