cytotox assays

KTano ktano at aol.com
Sat Jul 9 21:54:05 EST 1994


In article <1994Jul7.092654.67478 at kuhub.cc.ukans.edu>,
mejones at kuhub.cc.ukans.edu writes:

<We are trying LDH assays from Promega as an alternative to the
chromium release cytotoxicity assay. However, we have background
problems with the LDH assay. We are using J774 macrophages
stimulated with LPS. At >1 ng/ml LPS, we get high LDH readings
on the J774 cells alone.

Does anyone know of any other cytotoxicity asays (preferably
non-radioactive) that we could use? Or why we have these
background problems? (We don't have such high background
when we use bone marrow macrophages.) Any help would be
greatly appreciated.>

I have tried the Promega LDH assay but have not been impressed due to the
need for assessing the LDH content of the effectors themselves. Very often
(at least in my case) effectors are the limiting cells and I do not want
to use more than I strictly have to.
If you are using proliferating cells as the target you may want to try the
JAM test (Matzinger, P. 1991. The JAM test. A simple assay for DNA
fragmentation and cell death. J. Immunol. Methods  145:185). It is
radioactive but it is much easier than chromium release assays and the 
radioactive waste it produces is not too problematic (tritium).
Best of luck,
Caetano Reis e Sousa, D.Phil., National Institutes of Health, Bldg. 10, Rm
11D15, Bethesda, MD20892



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