Jim and Bret,
I would make the same recommendation to try Promega's Anti-ACTIVE Caspase-3
Ab.  It specifically recognizes a neo-epitope only present in the active
cleaved form. The antigen apparently is not present in the inactive zymogen.

There also are other antibodies against active caspase-3; but I do not think
they ae commercially available. One was developed (I think by by Kevin
Tomaselli's group) at IDUN Pharmaceuticals. It also recognizes a neo-epitope
on the active form of the enzyme. The third antibody I am aware of was
developed (I think by Don Nicholson's group) at Merck Frosst (perhaps Brett
is familiar with this one).  It was developed using crystal grade active
caspase-3. Apparenlty they had a few mg left over from their X-ray
diffraction studies. These have been reported to work in the literature.

Promega's antibody has been commercially available for ~1 year so there
probably are few published references; but you may be able to find some
information from their web site www.promega.com or perhaps their technical
services dudes which ususlly are excellent and very responsive.

Ty Lee
-----Original Message-----
From: Connolly, Brett M <brett_connolly@merck.com>
To: 'Jim Manavis' <jim.manavis@imvs.sa.gov.au>; 'HISTONET'
<histonet@pathology.swmed.edu>
Date: Wednesday, February 14, 2001 2:46 PM
Subject: RE: Caspase 3


Jim,

Caspase 3, like the other caspases exists in cells as a inactive proenzyme.
Cleavage (activation) of caspase 3 is the indicator for apoptosis. If
apoptosis is what you are looking at, then check out Promegas new
anti-active Caspase 3 antibody directed against the p18 cleavage fragemtn
(active form) of Caspase.

Brett


Brett M. Connolly, Ph.D.
Merck Research Laboratories
Department of Pharmacology
WP26A-3000
PO Box 4
West Point, PA 19486
Ph. 215-652-2501
FAX 215-652-2075
e-mail: brett_connolly@merck.com


> ----------
> From: Jim Manavis[SMTP:jim.manavis@imvs.sa.gov.au]
> Sent: Tuesday, February 13, 2001 12:56 AM
> To: Histonet
> Subject: Caspase 3
>
> What antibodies and retrievals are there being used for Caspase 3.
>
> Thanks
>
> Jim Manavis
> Senior Hospital Scientist
> Department of Neuropathology
> Institute of Medical & Veterinary Science
> Adelaide, SA, 5000
> Australia
> Phone: 61-08-8222-3668
> FAX: 61-08-8222 3204
> e.mail: jim.manavis@imvs.sa.gov.au
> Disclaimer: Not this little black duck!
>
>
>