Hi Dan,
For separating PBMC from monkey blood we use LSM (lymphocyte separation
medium) from ICN. For nemestrina monkeys we use 95% LSM (diluted with PBS)
and for fasciculari we use 90% LSM. Blood is diluted 1:1 with sterile PBS
and 20 ml is layered onto 15 ml LSM. Spin at room temperature at 400g for
30 min.
Fascicularis blood is quite messy, PBMC are always contaminated with
erythrocytes - hence we perform an additional step to lyse red blood cells
after LSM separation. Nemestrina usually gives clean cell separations -
with very little or no contamination.
I hope this helps.
regards,
Nadeem
Nadeem Sheikh,
****************************************************************************
************
RPRC Department of
Pharmaceutics
University of Washington University of Washington
Box 357331 Box 357610
Seattle, WA 98195 Seattle, WA 98195
voice : +1 206 221 2356
fax : +1 206 956 0573
e-mail : nadeem@bart.rprc.washington.edu
http://www.rprc.washington.edu/
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