Re: NH4Cl lysing

CINDY.JO.GROSS@monsanto.com
24 Sep 1997 08:51:11 -0500

Nancy,

We prepare a 10X ammonium chloride stock which is stored refrigerated
and prepare the 1X working solution from that; the working solution is
stored at room temperature, tightly capped, and is good for 1 week.

I've worked with rat blood and spleen samples and usually lyse my
cells before staining, but I know there are other procedures where
staining is performed first.

Cindy Gross, Monsanto Envir. Health Lab
e-mail: cjgros@ccmail.monsanto.com

______________________________ Reply Separator _________________________________
Subject: NH4Cl lysing
Author: owner-cyto-sendout@flowcyt.cyto.purdue.edu at INTERNET
Date: 9/23/97 6:36 PM


To all of you who uses ammonium chloride lysing on peripheral blood and
bone marrow samples for surface markers:
1. does the pH of the solution stay for more than 1 day? Ours goes from
pH 7.2 to as high as pH 7.7 on the next day. We have been making fresh
batch everyday.
2. do you stain first, then lyse or lyse first? Are there any
differences in fluorescence intensity with one method or the other? If
there is anything else we should be aware of in using nh4cl lysing vs
FACSLyse, ficoll etc. please advise. Thanks much in advance! Nancy Gin
- United Hospital - St.Paul, MN.