Re: lysing buffer
<< Previous Message | Next Message >>
From: | "Tony Henwood" <henwood@mail.one.net.au> |
To: | "'histonet'" <histonet@Pathology.swmed.edu>, "Berger, Jennifer" <jberger@lrri.org> |
Reply-To: | |
Date: | Fri, 9 Jul 1999 10:50:46 +0000 |
Content-Type: | text/plain; charset=US-ASCII |
Dear Jennifer,
> Does anyone have a recipe for or know where I can order Lysing Buffer?
Try the following technique:
Haemolysing haemorrhagic specimens causes fewer problems in
identifying individual cells (1). The removal of red blood
cells can be achieved using Ficoll-Hypaque gradient separation
or using a lysis solution such as isotonic ammonium chloride
(1):
SOLUTIONS:
1. Lysis solution
Ammonium Chloride 4.5g
Potassium carbonate 0.5g
EDTA 0.0186g
Distilled water 500mls
2. Hanks medium
METHOD:
1. Centrifuge bloody fluid.
2. Remove supernatant and add equal volume of lysis solution.
3. Resuspend and incubate for 5 minutes at 4oC.
4. Centrifuge, if blood still remains, then repeat from step
2.
5. Resuspend pellet in Hanks.
(1) Kuenen-Boumiester etal (1996) Acta Cytolog 40:475-479.
Tony Henwood
Senior Scientist
Anatomical Pathology
Royal Prince Alfred Hospital
Sydney, AUSTRALIA
http://www2.one.net.au/~henwood
http://www.pathsearch.com/homepages/TonyHenwood/default.html
<< Previous Message | Next Message >>