[Protein-analysis] Western blot using FACS cells

Melissa Mazan via proteins%40net.bio.net (by melissa.mazan from tufts.edu)
Thu Jan 3 12:19:11 EST 2008


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
508-887-4589



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