Promega Cytotox 96 assay

[Joan Redd]

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?