RE: apoptosis

From:"Connolly, Brett M" <>

Lee Ann,

I asked a similar question months ago and got no help other than a hearty
'good luck!' :-)

So I did some investigating and can offer two IHC suggestions:

Anti-activated Caspase-3.  Cleavage (activation) of procaspase-3 (CPP32,
apopain, YAMA) irreversibly commits the cell to the caspase cascade and cell
death via apoptosis. Antibodies are available that react only with the
Caspase-3 cleavage fragment(s) and not procaspase-3.

Similarly, anti-PARP p85 fragment. PARP (poly ADP-ribose) is the nuclear
substrate for activated Caspase-3 which cleaves PARP into the 25kDa and
85kDa fragments. Antibodies are available that react specifically with the
PARP 85kDa fragment.

Neither of these should stain necrotic cells as far as I know. As you point
out TUNEL labeling of fragmented DNA doesn't discriminate between the two

I wish someone would find a protein expressed only by necrotic cells.


Brett M. Connolly, Ph.D.
Merck Research Laboratories
Department of Pharmacology
PO Box 4
West Point, PA 19486
Ph. 215-652-2501
FAX 215-652-2075

> ----------
> From: 	Baldridge, Lee Ann[]
> Sent: 	Wednesday, November 08, 2000 8:12 AM
> To: 	'HistoNet Server'
> Subject: 	apoptosis
> Good Morning All,
> Does anyone have any ideas on a non-TUNEL staining procedure for
> apoptosis?
> I'm in need of a system that will not stain necrotic cells.  Thanks for
> any
> and all help.
> Lee Ann Baldridge
> IU Hosp.
> Indpls.,IN
> 317-274-5790
> 317-278-4820 fax

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