methods for cell sorting required
amcshea at world.std.com
Thu Apr 7 08:17:54 EST 1994
Ms S.P. Moyes (smoyes at crc.ac.uk) wrote:
: I would like to seperate antigen positive and antigen negative cell
: populations. Coating SRBCs or microtitre plates with the antibody has been
: suggested, then binding the cells followed by washing with PBS.
: Any comments on these methods or suggestions on other methods gratefully
: received .
Hmmm I have done a lot of cell sorting stuff, I'm not so sure about this
panning off the cells which dont stick method, it sounds a little hard
to control. How do you check for antigen +ve cells vs. antigen negative
cells? Are you using a simple cytospin or a FACscan? If you want to
separate small no.s find a FACs sorter and a friendly operator, if you
want to separate larger no.s oof cells, but sacrifice a bit of purity then
use magnetic sorting, the dynabead system is better for viability I hear
than the Miltenyi Biotech parmagnetic bead sytem that I used to use.
If you dont have access to this sort of equipment you should still be
able to enrich by panning but your purity might suffer somewhat.
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