keeping adherent cells ins suspension culture?

Wolfgang Schechinger wolfsc at ibms.sinica.edu.tw
Thu Apr 11 05:13:12 EST 2002


Dear all, 

How can I overcome this problem:

I have cells expressing an enzyme pinned on the extracellular part of the 
membrane. To assay this enzyme by FACS, I need to trypsinize the cells, but 
this (just 5 min) reduces the enzyme activity significantly. Detaching the 
cells with EDTA kills too many of them. 

What can I do to either 

1) obtain a single cell suspension or

2) to keep the cells for some time in suspension until some newly synthesized 
protein is delivered to the membrane? 

I welcome your input

Regards,

Wo
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Dr. Wolfgang Schechinger
Institute of Biomedical Sciences
Academia Sinica, Taipei, Taiwan

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