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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
much!!
Linda Weaver
Genetic Therapy, Inc.
Gaithersburg, MD email: weaverli@genetictherapy.com
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