anti-CD15 for L&L

Ian Dimmick Ian.Dimmick at newcastle.ac.uk
Tue Sep 26 15:59:27 EDT 2006


Dear Irina, only this week I have used CD15 both IgG and IgM clones with Ammonium
chloride(Pharmlyse) and they worked fine, with mature granulocytes and earlier
progenitors, you do not say what you mean by "damaged granulocytes" I assume you mean the
scatter pattern is not as you wish, but then if you use different lysing reagents ie
Ammonium chloride versus Facslyse tthen the scatter patterns will be slightly different .
If the scatter is very different then I would suggest you test your samples with just the
Ammonium chloride to see perhaps if this is the cause of your problem .

hope this helps 


Ian Dimmick
Flow Cytometry Core Facility Manager
Institute of Human Genetics
Bioscience Centre
Central Parkway
Newcastle Upon Tyne
NE1 3BZ
UK

Ian.Dimmick at ncl.ac.uk
Tel	0044 191 2418831/ 8825
Fax    0044 1912418666
(mob) 0044 7970344823



-----Original Message-----
From: Irina Grigorieva PhD [mailto:Irina.Grigoriena at Northside.com]
Sent: Mon 25-Sep-06 16:15
To: cyto-inbox
Subject: anti-CD15 for L&L
Hello Flowers,

I have a question about anti-CD15 antibodies for lymphoma/ leukemia immunophenotyping. 
Both  BD and Beckman-Coulter anti-CD15 Abs are damaging granulocytes, when an ammonium
chlorite lysing solution is used. When we use FACSLyse according to BD recommendations,
everything turns fine. But FACSLyse has so many other disadvantages for us, so we have
stopped using it and switched to ammonium chlorite a long time ago. Does anybody have any
solutions for this dilemma or any suggestions how to use anti-CD15 without using
FACSLyse?

Regards, Irina

Irina Grigorieva, PhD
Director, Flow Cytometry Laboratory
Northside Hospital, Atlanta, GA
(404)- 851-6541
e-mail: irina.grigorieva at northside.com








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