Re: CD14 and trypsin (and other receptors, too)

From: Dennis Broud 301-594-5879 FAX 301-594-3037 (BROUDD@CDER.FDA.GOV)
Date: Wed Jun 23 1999 - 10:15:50 EST

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Ditto what Julie Turner said

AND for human monocytes/macrophages you can lay the flask on the top of a bed
of wet ice after you add the EDTA in Buffer .  This will usually shorten the
time you need to get the cells to release from 15 minutes to 5-10 minutes.
(Seems to be more donor dependent than anything else.)  Julie is right, you
need to monitor the process microscopically to get consistent results.
Once the cells are loose,  add 5 to 10 volumes of media to dilute the EDTA
and spin the cells out, then don't be afraid to wash the cells with EDTA free
media and spin them out a couple of times.

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