LDH Assay Kit (Cytotoxicity) ab65393 uses WST for the fast and sensitive detection of LDH released from damaged cells.
The LDH assay, also known as LDH release assay, is a cell death / cytotoxicity assay used to assess the level of plasma membrane damage in a cell population. Lactate dehydrogenase (LDH) is a stable enzyme, present in all cell types, which is rapidly released into the cell culture medium upon damage of the plasma membrane. LDH is the most widely used marker used to run a cytotoxicity assay.
How the assay works
The LDH assay protocol is based on an enzymatic coupling reaction: LDH released from the cell oxidizes lactate to generate NADH, which then reacts with WST to generate a yellow color. The intensity of the generated color correlates directly with the number of lyzed cells.
Only 10μl of culture medium is required for the assay, and thus background from serum and culture medium is significantly reduced. Cells can be cultured in regular 10% serum containing medium; no reducing serum or special medium is required for the assay.
This LDH assay kit uses a chemical method to couple the NADH reaction with WST, which is more tolerant of variability in the sample than the diaphorase-based coupling method used in kits available from other vendors. The chemical method means that the active ingredients of the kit, excluding the LDH positive control, do not contain enzymes, meaning they are more stable for practical use in the lab in repeated experiments.
In addition, the WST used in the kit is highly stable, soluble in solution and unlikely to precipitate in solution, unlike the INT used in kits available from other vendors. This means that the reaction can be read multiple times over a long time-course. The reaction can also be stopped at any time point.
LDH assay protocol summary
- - Transfer 10μl culture medium into new plate
- - Add LDH reaction mix and incubate for 30 min at room temp
- - Analyze with microplate reader
LDH activity can be easily quantified by spectrophotometer or plate reader at OD450nm.
Related LDH assay products
If you would like to use a fluorometric assay, please refer to LDH-Cytotoxicity Assay Kit (Fluorometric) LDH-Cytotoxicity Assay Kit (Fluorometric)ab197004.
This kit is more sensitive than the colorimetric LDH Cytoxicity Assay Kit LDH Assay Kit (Cytotoxicity)ab65391.
To measure LDH activity in sample types such as serum, plasma, and cell lysates, we recommend LDH assay kit LDH Assay Kit / Lactate Dehydrogenase Assay Kit (Colorimetric)ab102526.
Other notes
This product is manufactured by BioVision, an Abcam company and was previously called K313 LDH-Cytotoxicity Colorimetric Assay Kit II. K313-500 is the same size as the 500 test size of ab65393.
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