From: Ian_DIMMICK@europe.bd.com
Date: Fri Jun 20 2003 - 16:26:03 EST
7-AAD will ingress into viable cells, this will happen at a variable rate
depndant on concentration of 7-AAD and temperature, as a rough guide I try
to analyse cells within 2 hours when using 7-AAD as a viability marker , if
you also make a concerted effort to ensure that care is taken as to what
type of lysing agent is used in sample preparation (although you did not
state whether this was part of your protocol) as some lysing agents will
allow 7-AAD to enter cells whether viable or not .
You say you ran the same samples , however I would be interested in
your PI results within the same tube
Ian
"Kristie Melnik" <kmelnik@imeddinc.com> on 19/06/2003 20:41:31
To: cyto-inbox
cc:
Subject: [7-AAD non-specific binding?]
Has anyone ever had experience with 7-AAD nonspecifically labeling viable
cells? I had cell sample preps analyzed yesterday on a FACSCalibur and
everything looked dead. When I ran the same samples with propidium
iodide, they looked fine. I also did a trypan blue exclusion just to
verify the viability of the cells. Both the PI and the trypan blue had
similar results. The funny thing is I used 7-AAD last week on the same
type of experiment and it worked perfectly. Anyone have any thoughts??
Thanks,
Kristie
Kristie M. Melnik
Associate Scientist
iMEDD Inc.
1381 Kinnear Rd, Suite 111
Columbus, OH 43212
Phone --614-340-6001
FAX-- 614-340-6005
kmelnik@imeddinc.com
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