Re: A GOOD monocyte marker?

Ronald L. Rabin, M.D. (rr84g@nih.gov)
Tue, 11 Mar 1997 08:09:30 +0800

>Hi,
>
>I have a rather simple sounding question, particularly for the sorters
>out there. I am looking to positively separate CD4, CD8, and monocytes
>from Ficoll-hypaque separated blood samples. I intend to use avidin
>columns and biotinylated antibodies. Obviously CD4 and CD8 will be use
>for the isolation of these lymphocytes, but the best monocyted marker is
>less clear. I have considered CD14, CD15, and CD33. My goal is to
>obtain a pure monocyte sample, not necessarily the entire monocyte
>population. The cells will be fixed immediately after they are
>isolated and the subsequent manipulations are not affected by the
>activation state of the cells. Does anyone have experience with these
>surface markers or others that would be more appropriate for isolating a
>relatively pure monocyte population?
>
>Thanks in advance
>
>Wallace Lauzon
>Dept. of Microbiology and Immunology
>University of Ottawa

100% of monocytes stain brightly with anti CD11c. Sigma sells a cy5-pe
conjugate, very good, or any of the PE conjugates are good. Some
lymphocytes do stain dim with CD11c, however, but discrimination on a
sorter is not a problem. Discrimination on a column would be a problem.

There are no specific markers for myelomonocytic cells analagous to
lymphoid cells. That is, though CD19 is B cell specific and CD3 is T cell
specific, CD14 eis expressed on neutrophils as well as monocytes, although
at lower levels. True for CD11c. Some markers, very bright, such as CD13,
are expressed on same levels on neutrophils and monocytes.

If you are looking for specificity, your best bets are CD14 or CD11c, both
of which are very bright, and can discriminate between the dull expressers,
neutrophils and less mature monocytes in the case of CD14, some lymphocytes
and neutrophils in the case of CD11c.

The last time I looked, CD33 wasn't that great, because it wasn't very
bright. CD15 is not best for monocytes becasue of the wide heterogeneity
of expression, and is much better for neutrophils, which are extremely
bright.

ron

Ronald L. Rabin, M.D.
Clinical Associate
Cytokine Biology Unit
LCI/NIAID NIH
phone: (301) 402-4910
fax: (301) 496-7383
e-mail: rr84g@nih.gov


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