In article <reddjo.14.0 at wwc.edu> reddjo at wwc.edu "Joan Redd" writes:
> We have been attempting to use the Promega Cytotox 96 colorimetric
> cytotoxicity assay for assaying the activity of rat CTL. In this assay we
> use AIM V serum-free media w/o phenol red from Gibco. The spontaneous and
> maximal LDH release of our target cells has been within expected
> parameters. However, the spontaneous LDH release of our effector cells has
> been extremely high, too high to calculate any meaningful % specific
> lysis. I would appreciate advice from anyone who has had experience with
> this assay. Which cell culture medium has worked the best? Has anyone
> successfully used this assay with rat CTL?
>
AIM V serum free medium is gastly stuff. It will not even support mitogen
driven responses by human PBMC. If you need a serum free environment then why
not try simple RPMI 1640? Also - very few systems beat good old 51Cr labeled
target cell release systems.
--
Another shitty day in paradise
Adrienne Morgan
