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how to use optimal antibody concentration

how to use optimal antibody concentration - Flow Cytometry Forum

how to use optimal antibody concentration - Flow Cytometry Forum. Discuss FACS cell sorting, flow cytometry, analysis and cell staining.


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Old 08-16-2008, 06:07 PM
Pipette Filler
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Default how to use optimal antibody concentration



Hi, All,

I want to analyze CD44 expression on peripheral CD8+ T cells from naive C57BL/6 mouse. but I found CD44 expression is heterogenous expression on CD8 T cells. Also, when I used saturated antibody to stain cells, the whole histogram peak moved to right with little overlap in isotype control. when I titrated down antibody concentration, the peak shifted to left. there is more overlap in isotype control. I do not know how to use correct antibody amount to stain the cells. In addition, how to define positive or negative expression? I feel that objective factors impact the data interpretation.

please help me to figure it out.

thank you

please reply email: [Only registered users see links. ]

I stained 10ul blood in 100ul volume.
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