Hello all, I'm trying to stain and isolate a rare population (1-2%) in bone marrow with PE-conjugated abs. I succeeded to label the cell for FACS using the following protocol: 1x10^6 nucleated BM cells/100ul FACS buffer with 2 ul Ab (1:50 dilution). The cells showed intermediate fluorescence. My question is: if I would like to stain 100x10^6 cells should I extrapolate the volumes so I get 100x10^6 MNCs/10ml FACS buffer with 200ul Ab??? I thought of keeping the Ab dilution but to increase the cells amount, lets say 100x10^6 cells in 1ml not 10ml, and in this case use 20ul of the Ab instead of 200ul. Please your advice. Best regards Hadi Aslan PhD, DMD Skeletal Biotech Lab/Prof. Dan Gazit The Hebrew University of Jerusalem- Hadassah Medical Center PO Box 12272, Jerusalem 91120 Israel Tel: 972-2-6757625 Fax: 972-2-6757628 Mobile: 972-545-909208 Web site: http://gazitlab.huji.ac.ilReceived on Wed Jun 25 12:38:00 2008
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