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CD 4 cell isolation

Mark Doherty mdoherty at pop.niaid.nih.gov
Thu Sep 4 10:05:22 EST 1997


In article <5uivgo$kgq$1 at nnrp4.snfc21.pbi.net>, "Scotty Wilcoxen"
<wilcoxen at hsc.usc.edu> wrote:

>  In our lab we often need to make T cell rich populations, specifically CD4+
> and CD8+, and we use the MACS system.  It gives great yields.  They use an
> anti-CD4 or anti-CD8 Ab conjugated to a metallic bead.  This solution is
> then passed through a column with steel shavings in it.  The column sits in
> a huge magnet, and your desired fraction is either eluted out, or remains in
> the column.   It is a bit slow, and VERY costly.  I hope this helps.

A useful trick if you are doing MACS separation for T cells is to do RBC
lysis and a nylon wool column first, followed by positive selection for
your population of interest. By cutting down the number of cells that goes
into the MACS analysis you greatly decrease the number of beads you use
(and thus cost) and also enhance your final purity.  By combining the two,
we routinely got CD4 yields in the high %90's range - almost as good as
FACs sorting and far faster, with better cell yields.  The nylon wool
columns are easy to make, easy to use and dirt cheap.

Cheers, Mark



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