Re: lysing buffer

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From:"Tony Henwood" <>
To:"'histonet'" <>, "Berger, Jennifer" <>
Date:Fri, 9 Jul 1999 10:50:46 +0000
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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.  Lysis solution
         Ammonium Chloride        4.5g
         Potassium carbonate      0.5g
         EDTA                     0.0186g
         Distilled water          500mls
2.  Hanks medium


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
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

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