NH4Cl Lysing

From: Vincent Gin (ssni@ix.netcom.com)
Date: Tue Sep 30 1997 - 12:41:53 EST


I have to say that all of you are terrific! Ask and you shall receive 
answers and support in all areas of flow cytometry! I sent out 
questions on NH4Cl lysing and received numerous answers the next day! 
I would like to clarify our search for the IDEAL lysing technique:
- it should NOT render live cells 7AAD positive, since we use 7AAD as 
the viability probe (therefore FACSLyse, F&P etc. is out).
- easy and cheap to make, quality maintained controlled. 
To measure out the reagents, pH each day and still confront variation 
in lysing capability is NOT acceptable. I am also very concerned about 
the fact that there were reports - Cytometry Vol 30, No.3 June 15, 1997 
(also mentioned in Barry Grimes' reply) that there are unexplained 
abnormal decrease in CD4 expression using NH4Cl lysing. Has anyone of 
you noticed this phenomenon and how do you address this? 
By the way, our recipe include: 0.829 g NH4Cl, 0.100 g KHCO3, .0037 g 
EDTA per 100mL, pH 7.2 @ RT. Thanks much for all your help. Nancy Gin - 
United Hospital, St.Paul, MN.  



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