Re: neutrophil/monocyte/platelet staining

From: Bruce Davis (DAVISB@mmc.org)
Date: Thu Sep 13 2001 - 07:20:04 EST


In what sense are you studying activation?  If it is in vivo activation, I recommend CD64
(both monocyte and PMN upregulate secondary to INF-gamma, G-CSF and other cytokines)
and CD163 (modulated on monocytes by glucocorticoids and cytokines).


Regards,



Bruce H. Davis, M.D.
Maine Medical Center Research Institute
81 Research Drive
Scarborough, Maine  04074
USA

PHONE: 207-885-8113
FAX: 207-885-8110
Email:  davisb@mmc.org

>>> Rekha Bansal <bansalr@Gliatech.com> 09/12/01 09:30AM >>>

Hi,
I am new in this field and interested in studying neutrophil, monocyte, and
platelet activation.  To prepare and stain cells what do you think would be
a good method of choice.  Also would like to know what CD markers are
specific to each of these cell types.  For neutrophils I am using
CD15/CD11b, for monocytes I am using CD14/CD11b, and for platelets I am
using CD42a and CD62P.  Are there more specific CD markers available?.

I do staining in PBS,1%BSA, 0.1% sodium azide, 20 mM EDTA.  (I added EDTA to
prevent further complement activation).  Would EDTA in anyway compromise the
staining?

What G forces are optimal for each of the cell types: I am currently using
the following neutrophils/monocyte (300 or 500 g), platelets (1200 g).

Rekha Bansal, Ph.D.



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