Re: caspase-3

From: Candace Enockson (enockson@musc.edu)
Date: Wed Sep 08 1999 - 14:42:35 EST


I have a client who has used this in virally infected cells with some
success.  However,there is not very good separation between negative and
positive.  I was told by a Pharmingen person that this was a problem with
this particular lot of antibody. Also, it does have a lot of non-specific
background over non-stained cells. Hopefully they will be coming out with
the monoclonal version soon.

Candace Enockson
Medical University of South Carolina


--On Tue, Sep 7, 1999 9:35 AM -0400 Maryalice Stetler-Stevenson
<stetler@box-s.nih.gov> wrote:

> 
> We also have tried this antibody in parallel with Annexin V without
> success but haven't done a time course, etc to determine if there are
> biological reasons this isn't working for us. I would be interested in
> others experience with this antibody and any tricks people may have used
> to get it to work better.
> 
>	 Maryalice
> 
>> Dear all
>> 
>> We have been using a anti-Caspase-3 rabbit polyclonal ab conjugated
>> with PE (this is supposed to bind to the active form only) by flow.
>> We've looked for caspase-3 activation in camptothecin or etoposide
>> treated Jurkatt cells (cytospins confirm apoptosis in these
>> cells) after fixation and perm, without success. This ab is from
>> Pharmingen.
>> 
>> Does anyone have experience with looking for caspase-3 activation by
>> flow ? Are there other antibodies available ?
>> 
>> Any comments would be much appreciated.
>> 
>> Fran Gibson
>> Dept Haematology
>> St. George's Hospital Medical School
>> London
> 
> Maryalice Stetler-Stevenson
> Director Flow Cytometry Unit
> Laboratory of Pathology, NCI, NIH



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