fluorogenic substrate for caspase 3 -Reply

From: David Hedley (david_hedley@pmh.toronto.on.ca)
Date: Wed Jan 28 1998 - 08:18:41 EST


Padma writes:

"Has anybody out there  tried using fluorogenic substrates to measure
caspase 3 activity during apoptosis by flow.
Any help with info on reagents and vendors will be greatly appreciated."
____________________________________

We had a go using the Biomol AMCA-DEVD fluorigenic substrate for
caspase 3 (aka CPP-32/yama).  It gave a great fluorescence signal, but it
all washed out of the cells.  In their catalogue Biomol mention that they are
working on a probe that yields a product with better cell retention. I think
this would be a very useful reagent if it could be made to work.  It's time
we had some reagents for flow enzymology that measure things of real
biological interest!

David Hedley
Ontario Cancer Institute
Toronto



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