In article <5fnpll$mbd at news.bu.edu>, Robin Blumenthal <robinb at bu.edu> wrote:
>I am interested in using magnetic beads to seperate CD4+ T cells from PBMCs.
>Has anyone else tried this and what types of purification were you able to
>obtain? Did you do any other steps (such as treat with complement) to
>increase purity?
>>Thanks!
>>--
>Robin Blumenthal
>robinb at bu.edu>(617) 638-5218
>
I have used Dynal beads before and get pathetic purity around
80-90%, but that is starting with a relatively clean population. Some in
the lab use the Miltenyi Biotech column system with very good results (~98+%
pure). The drawback is that the colums and magnetic supplies are costly.
The initial setup is around $2000. For most work, I use either SRBC or nylon
wool columns to get a fairly pure T cell poplulation, then I pan against
CD8 and CD16 (NK's). I usually get >95% CD4 T cells. If you use nylon wool,
you might need to add a few other mAbs to your panning step since some other
cell type do get through. This is for human cells, it may be different for
mouse or other species.
Good Luck,
--
Raymond Doty
Dept. of Immunology
University of Washington
Seattle, Washington
