CD 4 cell isolation

Mark Doherty mdoherty at pop.niaid.nih.gov
Tue Sep 9 09:09:02 EST 1997


In article <3414cab5.525757 at news.oz.net>, Mark@(NO SPAM)mizuno.com (Mizuno)
wrote:

> Geez.... I remember the old days when I used to boil the nylon wool in
> acid, wash it, dry it, and then tease it, before weighing out the
> proper amount and shoving it into a 20ml or larger syringe -- followed
> by autoclaving the damn thing! (Am I showing my age here?!!  ;)

No, Not really.  I remeber doing all that too.  It's simply that these days
you can buy nylon wool already cleaned up.  I like to give the stuff a
quick boil (toss it in the microwave in a 500 ml beaker for 6 minutes, then
empty it out on some foil and leave to dry overnight in the incubator) and
then remove any lumps (2 minutes with a carding comb makes enough for a
column).  I find that an hour devoted to making columns on a slow day and
autoclaving them gives me a stack of 20 or so that I can use at will on
busy days (such as pretty much any day when T cell purification is the
order of business!) - and it's a time saver because when the cells are on
the column I can wash the MACS beads, get my purification set up ready for
staining - and then when ready simply wash the cells out of the nylon wool
and bingo! You're ready to roll.  Personal preference, I guess.

I do agree with your other comments on the relative merits of Dynabeads and
MACS though.  The extra cost of MACS is worth it.  Dynabeads do seem to
work OK for doing negative depletions, and are a cheaper alternative if you
do a lot of those or to selectively clean up a population before MACS or
FACS seperation.

Cheers, Mark



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