IUBio

MACS FACS Dynabeads

Sebastian Bunka bunka at i112pc09.vu-wien.ac.at
Fri May 29 02:55:28 EST 1998


Hi all,

we are on the way to analyze antigen specific T-cell proliferation in CD4+
cells and are looking for ways to isolate them.

I know that several people are using the MACS columns (Miltenyi) with
good results. Any opinions that the magnetic beads (they are rather
small and do not influence FACS analysis) have influence on
proliferation assays.

I talked now with a guy offering Dynabeads. But they are huge
(I think more than 5 micrometer). They HAVE to be stripped off.
Does anyone has experience in this?

And what about FACS sorting. We can use this small new BD FACSscan
equipped with a sorting extension. Do one get enough cells
for proliferation assay (we are using this stuff based on BrdU
in microtiter plates - Boehringer Kit). Sure want to run
in triplicate and with controls and several peptides of a protein.
And, are the cells viable enough? Can the be frozen after FACS?

Any comment greatly appreciated! Thanks in advance!

Sebastian

-- 
Sebastian Bunka, Dr. med. vet
Inst. Med. Chemistry, Vet. University Vienna
Ph. +43-1-250 77 ext. 4208, Fax: ext. 4290
e-mail: Sebastian.Bunka at vu-wien.ac.at



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