From: Ian_DIMMICK@europe.bd.com
Date: Wed Apr 10 2002 - 02:47:45 EST
I have used 7-AAD for some time now for this application and I find the following rules very helpful 1 Remember that 7-AAD is also capable of being absorbed into live as well as dead cells , the uptake into dead is rapid , but into live much slower , however to get maximal differentiation from live to dead analyse as soon as you can after the initial antibody incubation period is complete 2 Be careful which Lysis system you use, some will permeabilise slightly and consequently can not be used with this dye, try to use an ammonium chloride based lysis 3 7-AAD is quite bright to the point it may start to quench some detectors , do not apply ver y high volts/gains , it is not necessary and may cause you to have to run with high compensation needlessly 4 I use 5ul of 7-AAD in with the 45/34 cocktail , this works well (using 7-AAD from BD-Pharmingen) Hope this is helpful Ian
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