Hi all -
I am trying to identify proteins using WB on cells that have already
been through FACs. We make a single cell suspension from lung, bring it
on ice to our core facility (about 45 minutes away) and then bring the
sorted cells back to the lab in media on ice. About 4-5 hours in all.
The sorted cells I receive range from 1200 cells/ 250 ul to 1x10 to the
7th cells. I wash the cells at slow speed in PBS (4C) when I receive
them, then extract protein using M-Per from Pierce. I'm not having much
luck with samples that have less than a million cells. Any suggestions?
Many thanks - Melissa
Melissa R. Mazan, DVM, Diplomate ACVIM
Associate Professor and Director of Sports Medicine
Tufts Cummings School of Veterinary Medicine
200 Westboro Road
North Grafton, MA 01536