methods for cell sorting required

Andy 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.

	Good luck,

			Andy.




More information about the Methods mailing list