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CTL assay - CTL assay question (Nov/02/2010 )

I am trying to use the LDH based CTL assay kit from Roche. I'm using mouse tumor cells as my targets for lysis with C57 splenocytes primed in vitro by the same irradiated cells for 5 days. My problem is I'm getting negative values as the spontaneous release is so high.
I've been using complete media for 4 hr incubation of effector:target.
Please help. We don't have radioactivity permit so I cant use CR51 release assay.

-tubs-

Time to try another method. Here's one example based on flow cytometry.

http://immunochemistry.net/Cytotoxicity.htm

Best and let us know.

-CellSpecific.com-