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I've found it easier to use the commercial kits from
Orpegen/Germany (or BD) as neither the extracellular FITC
quenching by EB nor by low pH seems to be satisfactiory. The
kit gives us very consistent / reproducible results
regarding both, phagocytosis and burst worthwhile the price.
Gerhard Nebe-v.Caron
Unilever Research, Colworth,
Sharnbrook, Bedfordshire
GB - MK44 1LQ
Tel: +44(0)1234-222066
FAX: +44(0)1234-222344
gerhard.nebe-von-caron@unilever.com
______________________________ Reply Separator _________________________________
Subject: Coulter Immuno Lyse
Author: pmcclosk@libserv1.ncmc.co.nassau.ny.us at INTERNET
Date: 26/11/96 07:59
Our laboratory is using FITC labeled bacteria to study phagocytosis in
neutrophils. Fluorescence of both the extracellular and adherent
bacteria is quenched with ethidium bromide. In the future, we would like
to simultaneously measure oxidative burst in neutrophils using DCFH-DA
and HE.
Recently, we came across a reference where Coulter's Immuno lyse was
used to quench extracellular FITC labeled bacteria with great success.
We tried this method and were unsuccessful in our attempts to quench
extracellular bacteria.
Has anyone tried to use this method for quenching? And it so, were you
successful? Any suggestions would be appreciated.
Thank you.
Pamela McCloskey
Research Scientist II
Department of Pediatrics
Nassau County Medical Center
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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
Web