4.2.2 (b) Lactate Dehydrogenase (LDH) release assay to identify
cell cytotoxicity
Lactate dehydrogenase is a cytosolic enzyme which is released into the
cell culture media with plasma membrane damage which can be measured.
Levels of LDH released in the media are an indicator whether the cells
are undergoing necrosis. High levels of LDH in the media are
proportional to cell death via necrosis. Plates with unstimulated cells
and Rv2615c or control (LPS/Staurosporine) stimulated cells were
centrifuged and 20µl of media from each well and 20µl reaction buffer
were added to fresh wells. Plates were incubated for 30 min at room
temperature following addition of 20µl stop solution. Absorbance reading
was recorded at 490nm.