The Good
Simple to use and fast results
The Bad
Stability of WST substrate Mix is not as described in the kit protocol. For best results, only use substrate mix kept at 4C up to 1-2 weeks.
The Bottom Line
The LDH assay assesses the level of Lactate dehydrogenase (LDH) released into the cell culture medium upon damage of the plasma membrane. LDH released from the cell oxidizes lactate to generate NADH, which then reacts with WST (from the Kit) to generate a yellow color. The absorbance (OD 450nm) of the generated color is directly related to the number of lysed/damaged cells.
It is useful for drug screening as it is a rapid and simple assay, which allows assessment during continuous experiments. Cells can be kept in culture after assay.
To take best advantage of this assay, one must optimize absorbance reading times to the specific cells and use the correct controls. A negative control comprised of cells incubated with only medium and a positive control comprised of cells incubated with medium with 10%Lysis buffer (provided in the kit).