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LDH-Cytotoxicity Assay Kit II (ab65393) is based on using WST reagent for a fast and more sensitive detection of LDH released from damaged cells. The assay utilizing an enzymatic coupling reaction: LDH oxidizes lactate to generate NADH, which then reacts with WST to generate yellow color. The intensity of the generated color correlates directly with the cell number lysed. Since WST is brighter than other viability reagent, less amount of culture medium is required for the assay, and thus the 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. In addition, since the WST is very stable, the reaction can be read multiple times and can be stopped at any time point during the reaction. LDH activity can be easily quantified by spectrophotometer or plate reader at OD450nm. The kit provides all necessary reagents including LDH positive control.
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Cell death or cytotoxicity is classically evaluated by the quantification of plasma membrane damage. Lactate dehydragenase (LDH) is a stable enzyme, present in all cell types, and rapidly released into the cell culture medium upon damage of the plasma membrane. LDH, therefore, is the most widely used marker in cytotoxicity study.
If you would like to use a fluorometric reading, please refer to LDH-Cytotoxicity Assay Kit (Fluorometric) (ab197004).
|Cell Lysis Solution||Clear||1 x 5ml|
|LDH (lyophilized)||Red||1 x 1µl|
|LDH Assay Buffer||NM||1 x 50ml|
|Stop Solution||1 x 5ml|
|WST Substrate Mix||Amber||1 vial|
Our Abpromise guarantee covers the use of ab65393 in the following tested applications.
|Functional Studies||Use at an assay dependent dilution.|
Bafilomycin A1 (BafA1) toxicity was assessed in Human hepatic (Huh7.5) cells after 24 hours at different concentrations (12.5nm, 25nm, 50nm and 100nm) were administered to cells using LDH cytotoxicity assay kit (ab65393). Cytotoxicity was measured by subtracting LDH content in remaining viable cells from total LDH in untreated controls.