Rat spleen cells

roberta rigolio roberta.rigolio at gmail.com
Wed Jan 31 10:10:02 EST 2007


Hi everyone,

I'm a naive flower. I would like to work on rat spleen cells to investigate
the level of the adhesion molecules expression (LFA-1 and VLA-4) by flow
cytometry.

I am trying to obtain single cell suspension free of red blood cells using
density gradient and a mild treatment with NH4Cl to the leukocyte collected
at interface.
Do you think this method (25 min centrifuge on density gradient+ NH4Cl) can
influence the level of adeshion molecules on the leukocyte surface?

I have tried also with the osmotic lysis using water, without good results.
Have you got some useful protocol to suggest me on rat (tpe of water and
cell solution /water ratio?

Thanks a lot for all your help.
Roberta




-- 
PhD in Neuroscience
Dept. Neuroscience and Biomedical Technologies
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20052 Monza
MI - Italy
Phone: +39 02 64488123
Fax: +39 02 64488253
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