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We routinely do FACS analysis of CHO cells, looking at cell surface
receptors with peptides and antibodies, as well as internal receptors.
We grow the cells on plastic, glass, laminin, fibronectin, etc. We
remove them with an incubation with calcium and magnesium free medium,
2x, about 10 min per incubation, then a brief exposure to Trypsin, before
rinsing the cells off the substrate with culture medium. After removal
we place on ice, wash 2x with PBS before labelling. Protocols for
labeling vary depending on label, external or internal. The most
important part of the prep is the use of Calcium and Magnesium free
medium, and filtering the cells to get rid of clumps just prior to
analysis, which ensures single cell suspensions. Let me know if you
want more information.
Deb Berglund
Montana State University]
Microbiology
On Fri, 23 Aug 1996, Martin, Jill V. wrote:
>
> We are studying the expression of a protein with several putative membrane
> spanning domains using synthetic peptide antibodies. We are interested in
> hearing from individuals with experience in FACS analysis of substrate
> attached CHO cells, including methods of detachment, suspension,
> permeabilization and fixation suitable for FACS analysis.
>
> You may reply to Jill Martin as above, or to me directly:
> mcdonald.john@mayo.edu.
>
> Thanks in advance, John McDonald, Mayo Clinic Scottsdale.
>
>
>
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