Does increase LDH activity induced by stress (toxicants) in cell culture medium indicate cell necrosis ??
LDH activity can be used to detect cytotoxicity (e.g. the kit from Roche)
But you have to pay attention to some special points:
1. do not freeze the probes
2. to calculate cytotoxicity the measurement of medium LDH and cellular LDH ist advisable.
3. always use controls with untreated cells, medium control, low and high control (see manual)
4. Calculations:
Cytotoxicity [%] = (extracellular LDH from medium)/ (cellular LDH + extracellular LDH)
Viability [%]: (cellular LDH)/(cellular LDH + extracellular LDH)
Best Regards
Anne-Helen Lutter
Dear Anne-Helen Lutter,
Thank you for nice and specific explanation of LDH assay method. Leakage of LDH in the medium certainly an evidence of cell death as well as cytotoxicity. My concern is whether the death is necrotic or other (e.g. apoptosis). To explain the cell death process the LDH activity together with other factors like Cas9/Cas3 may be a clear indicator.
Thank you for your nice and helpful answer.
Necrosis is associated with a Plasma membrane leakage where intracellular contents is released into the extracellular milieu. This process can be measured with LDH activity
see article: doi:10.1007/978-1-62703-290-2_7
I would first check for apotosis with the Apoptosis/ Necrosis Detection Kit (blue, red, green) from abcam (ab176750) or with other factors to detect apoptosis. If there is no apoptosis then LDH measurement will indicate necrosis.
Hi everyone,
Can i use LDH assay to detect cell membrane leakage of bacterial cells?
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