Promega Cytotox kit
elliot at news.acns.nwu.edu
Tue Jul 19 16:22:02 EST 1994
I spoke with Promega's tech-help. They have heard many cases where primary (
freshly explanted) splenocytes showed high spontaneous LDH release.
Their suggestions were to decrease the time of incubation (effectors vs
targets in vitro) and decrease the time of substrate conversion by the
enzyme (dark incubation of supernatants and substrate mix). I've tried this
with some success. I'm measuring CTL response between cells with MHC
mutations. I don't expect a whopping response because of the minor
differences, and I'm not certain that the slight response I see with this
kit is due to CTLs or assay-difficulties.
If anyone knows if this or any other nonradioactive CTL protocol (Molecular
Probes has a live-dead fluorescent kit) that is reliable for small
differences in response, please let me know!
Thanks in advance.
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