Simultaneous FDG and surface markers

Weaverli weaverli at
Mon Mar 25 10:18:00 EST 1996


Has anyone had substantial experience staining the same population of 
cells for B-galactosidase activity using FDG AND cell surface proteins 
using direct or indirect detection?  

IF you stain for the surface marker first (ON ICE, of course), then 
load the cells with FDG for 1 min, do you run the risk of experiencing 
down-regulation of the surface marker, or is 1 minute short enough 
that the cells still retain decent surface staining?

That hypotonic FDG loading sometimes has a tendency to render the 
cells quite fragile.  My inclination in the past was to refrain from 
excessive centrifugation and resuspension post-FDG loading.  
Therefore, my instincts tell me that surface staining after hypotonic 
loading should probably not be considered.

Am I on the right track?  Does "surface stain first, FDG-load second" 
sound like a reasonable strategy?

If you care to elaborate further on this, I welcome any other 
advice/considerations I have not mentioned in this post!  Thanks very 

Linda Weaver
Genetic Therapy, Inc.
Gaithersburg, MD      email: weaverli at

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