Promega Cytotox 96 assay

Adrienne Morgan Adrienne at dumpling.demon.co.uk
Wed Jul 20 13:47:14 EST 1994


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



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