RE: anti-caspase antibodies

Barbara Maloney wrote:

<how does it work with other organisms? e.g marine invertebrates?>

Hello Barbara,
We successfully detected caspase-3 activation using this Ab in several
human cell lines as well as on normal human lymphocytes. We did not
study, however, marine invertebrates. However the apoptotic machinery,
including caspases, is highly conserved through evolution. I would
expect, therefore, that this Ab may be useful to detect caspase-3
activation in different species.

In response to the "anti-caspase antibodies" query Alex Spurmanis wrote
that FLICAs may be used to detect activation of these proteases. Indeed,
FLICAs are very useful reagents to detect apoptosis and caspase
activation. Their virtue is simplicity in use - the cells are incubated
in their presence, then rinsed and their fluorescence measured by
cytometry. However, one has to be careful with data interpretation
because there may be a non-specific (non-caspase) component in their
binding to apoptotic cells (see Pozarowski et al.,Interactions of
fluorochrome-labeled caspase inhibitors with apoptotic cells. A caution
in data interpretation. Cytometry 55A: 50-60, 2003).

Zbigniew Darzynkiewicz, M.D., Ph.D.
Brander Cancer Research Institute at NYMC
19 Bradhurst Avenue, Suite 2400
Hawthorne, N.Y. 10532
darzynk@nymc.edu

http://www.darzynkiewicz.com/zbigniew/




-----Original Message-----
From: barbara maloney [mailto:maloneyb@fiu.edu] 
Sent: Thursday, July 07, 2005 9:26 PM
To: cyto-inbox
Subject: Re: anti-caspase antibodies

Darzynkiewicz, Zbigniew wrote:

>Frances Gibson writes:
><Am interested in looking at caspase activation by flow. 
>Looked into this a few years ago when doing westerns for caspase 
>activation, and didn't seem to be monoclonals fluorochrome confugated 
>for flow that were available and used and data published.
>Does anyone have experience, information, references ? Any feedback 
>would be great>
>
>We have extensive experience with the use of activated (cleaved)
>caspase-3 Ab. The polyclonal rabbit anti-caspase-3 Ab available from
>Cell Signaling works very well. We used it both, in conventional assays
>as well as in conjunction with ZENON  Molecular Probes technology. E.g.
>see:  Pozarowski et al., Interactions of fluorochrome-labeled caspase
>inhibitors with apoptotic cells. A caution in data interpretation.
>Cytometry 55A: 50-60, 2003. 
>See also Huang et al at:
>http://www3.interscience.wiley.com/cgi-bin/fulltext/107632598/PDFSTART
>
>
>Zbigniew Darzynkiewicz, M.D., Ph.D.
>Brander Cancer Research Institute at NYMC
>19 Bradhurst Avenue, Suite 2400
>Hawthorne, N.Y. 10532
>darzynk@nymc.edu
>http://www.darzynkiewicz.com/zbigniew/
>
>how does it work with other organisms? e.g marine invertebrates?
>  
>
Thanks
Barbara

>-----Original Message-----
>From: Frances Gibson [mailto:fgibson@sgul.ac.uk] 
>Sent: Wednesday, July 06, 2005 4:23 AM
>To: cyto-inbox
>Subject: anti-caspase antibodies
>
>  
>