Heather,
It's been years since I did this sort of thing, but I wonder if the 
temperature has more to do with cell functions such as patching and 
capping than with Ab-Ag binding efficiency.  You don't mention whether the 
cells were fixed, or whether you are looking at surface or intracellular 
Ag.  I my past life I once did experiments looking at surface Ag's on 
lymphocites that were unfixed.  We kept everything on ice to inhibit 
patching and capping until the experiment reached the point where the 
cells were fixed.  Can't remember the details, unfortunately.

Good luck with your experiments,
Karen Zaruba, Advanced Biologist
3M Company, St. Paul, MN 55144 
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Subject:        temperature of facs preps



Hi FLOWers,

About 5 years ago when I was analysing lymphocytes, we used to have the 
cell preparations at 4°C for labelling with the antibody.
The work I do now on whole blood, the cells are labelled at room 
temperature.

Why do people label at 4°C when it works perfectly well at room temp?

Heather

Heather Medbury (PhD)
Department of Surgery
Westmead Hospital
Westmead 2145
61-2 9845 7677
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     All experiments are preliminary

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  "In his heart a man plans his course
    but the Lord determines his steps"

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