Caspase/Apoptosis detection in non fixed cells

[Benoit Labarthe]

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Good morning!

I'm looking for an apoptosis labelling for non fixed cells, ideally this staining would not stain the cytoplasma nor the plasma membrane. I found the Nucview Caspase detection kit that detect caspase activation but stains the nucleus and seems to be. Does someone have experience with this product or an alternative solution (we will also use the PI exclusion staining)?

Thanks a lot!

benoit

--
Benoît Labarthe, PharmD, MSc
Institut de Cardiologie de Montréal
Montreal Heart Institute
Centre de Recherche, Laboratoire du Dr.Théroux
5000 Bélanger Est
Montréal Québec Canada H1T 1C8
Tel [phone] ext 3017
Fax (514)- 376 1076

[Sarah Aubut]

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http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Hello,

I have been using a kit from Immunochemistry technologies, LLC, it is the
Poly-Caspases FLICA kit, and is supposed to detect active caspases in fixed
cells, but I have also tried using it in living cells. Unfortunately, I have
not been able to develop a positive control. I'm working with a spruce
budworm cells line, Cf203, and I have had trouble inducing apoptosis that is
detected by this kit. It seems that most people who have published data
through the use of this kit have used FACS, and not fluorescence microscopy.
So in other words, I haven't found a successful way to detect caspases!
I have also tried detecting apoptosis through the use of a phosphatidyl
serine antibody, and have had some success here. This I have only done in
fixed cells.

I would be interested in any other information about possible methods of
detecting apoptosis!

Sarah Aubut
Cell Biologist, Microscopy Suite
Canadian Forestry Service
Sault Ste Marie, ON
Canada

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