Simple Apoptosis Protocol Requested

David Voehringer ylek at utmdacc.uth.tmc.edu
Wed Jul 12 18:45:23 EST 1995


In article <3u07ql$k5d at mailer.york.ac.uk> Dr Graham Timmins <gst3 at york.ac.uk> writes:
>From: Dr Graham Timmins <gst3 at york.ac.uk>
>Subject: Simple Apoptosis Protocol Requested
>Date: 12 Jul 1995 10:23:17 GMT

>I would be grateful to anyone who could point me in the direction of an easy and 
>reliable DNA fragmentation assay for the determination of apoptosis (in murine 
>primary cell lines if poss.) Thanks. gst3 at york.ac.uk

There are many assays that will measure cell death (MTT, trypan blue 
exclusion, etc..) based on membrane integrity or metabolic potential.  
Assays that measure cell death as a function of DNA fragmentation are based  
either on insitu end labeling (TUNEL) of the fragments or sorting of the 
intact DNA from the fragmentated DNA and quantifying the DNA using the 
diphenylamine method of Burton (Burton, K. 1956. Biochem. J. 62:315).  The 
second method is more cost effective and can be more reliable.  A good 
reference is, Sellins, K. and Cohen, J.J., J of Immunology, vol.139, No. 10, 
pp 3199-3206,  as well as our own, Story, M.D., Voehringer, D.W., Malone, 
C.G., Meyn, R.E., Int. J. Radiat. Biol., 1994, vol 66, no 6, 659-668.  This 
method works well on suspension cells with a little modification for adherant 
lines.  Email me if you need more help.
David W. Voehringer
Department of Experimental Radiotherapy
Univ. of Texas M.D. Anderson Cancer Center
1515 Holcombe Bvld.
Houston, Tx. 77030
(713) 792-3797



More information about the Cellbiol mailing list