RE: depletion of platelets

From: Keith Bahjat (KeithB@Cytomation.com)
Date: Wed Nov 21 2001 - 19:06:34 EST


Christiane,

I have a couple of suggestions:

First, we typically wash our PBMCs repeatedly with HBSS+1%BSA, spinning at
around 220 xg. We repeat these washes until the supernatant is clear. Time
consuming, but effective.

A new technique you might try utilizes products from StemCell Technolgies in
Vancouver. It's a product line called RosetteSep. They use antibody dimers,
where the tail of the dimer is an anti-glycophorin antibody, to cause
unwanted cells to aggregate and pass through the Ficoll gradient. I think
they include an anti-platelet antibody in their product list, but if not,
they can make you a custom cocktail for a reasonable price. If you want
quick results and cost is not an object, give it a shot.

http://www.stemcell.com/

Best of luck.

kb

p.s. Of course, I would NEVER suggest you use anything for cell separation
besides a MoFlo.........  :-)  Happy holidays.


------

Keith Bahjat, Ph.D.
Applications Engineering Manager
Cytomation, Inc
Fort Collins, Colorado
Phone: (970) 226-2200 x223
Fax: (970) 226-0107
keithb@cytomation.com



-----Original Message-----
From: Dr. Christiane Hollmann [mailto:ch@adnagen.com]
Sent: Wednesday, November 21, 2001 1:51 AM
To: cyto-inbox
Subject: depletion of platelets



Dear Flowers,
I have a problem to get rid of remaining platelets after MNC preparation via
Ficoll gradient. Up to now we centrifuge the MNC through FCS or BSA. Are
their any protocols to increase the efficiency of platelet depletion? Please
let me know!
Regards
Christiane




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