LDH-Glo™ Cytotoxicity Assay Technical Manual
Literature # TM548
Lactate dehydrogenase (LDH) is a soluble stable cytosolic enzyme present in many cell types that is rapidly released into the cell culture medium upon disruption of the plasma membrane. LDH is a widely used marker in cytotoxicity studies.
The LDH-Glo™ Cytotoxicity Assay provides a simple bioluminescent method for quantifying LDH release. The bright luminescent signal provides the sensitivity to determine cytotoxicity in samples low in cell number such as 3D microtissue spheroids, microfluidic cell culture chips, primary cells and stem cells.
In the LDH-Glo™ Assay protocol, LDH Detection Reagent (containing Lactate, NAD+, Reductase, Reductase Substrate and Ultra-Glo™ rLuciferase) is added to a sample of diluted cell culture media. If the sample contains LDH, the enzyme-coupled reactions start and progress simultaneously. The luminescent signal generated is proportional to the amount of LDH in the sample. The signal increases until all reductase substrate is consumed and the reaction signal is no longer in the linear range of the assay.