Coulter Immuno Lyse

Gerhard Nebe-von-Caron Gerhard.Nebe-von-Caron at unilever.com
Tue Nov 26 12:58:12 EST 1996



          Hi Pamela

          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 at unilever.com


______________________________ Reply Separator _________________________________
Subject: Coulter Immuno Lyse
Author:  pmcclosk at 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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