IMPROVED PROTOCOL FOR COLORIMETRIC DETECTION OF COMPLEMENT-MEDIATED CYTOTOXICITY BASED ON THE MEASUREMENT OF CYTOPLASMIC LACTATE-DEHYDROGENASE ACTIVITY
A. Sepp et al., IMPROVED PROTOCOL FOR COLORIMETRIC DETECTION OF COMPLEMENT-MEDIATED CYTOTOXICITY BASED ON THE MEASUREMENT OF CYTOPLASMIC LACTATE-DEHYDROGENASE ACTIVITY, Journal of immunological methods, 196(2), 1996, pp. 175-180
The conventional cytotoxicity detection protocols based on the lactate
dehydrogenase (LDH) activity assays rely upon the quantitation of the
enzyme activity released into the assay medium upon cell lysis. In th
e case of complement-mediated cytotoxicity this results in the need to
take into account the high and variable background LDH activity from
the serum added to the cells. Using primate-derived COS-7 and pig kidn
ey PK15 cell lines we show that, in the case of adherent cells, it is
possible to overcome this drawback and to measure cell death caused by
complement attack, by quantifying the amount of LDH activity retained
by the undamaged cells. The modified assay is therefore quicker to ca
rry out than the conventional procedure and cheaper because fewer cont
rol readings must be taken.