Detection PrincipleLactate dehydrogenase(LDH) is an enzyme that presents in almost cell types and it catalyzes the oxidation of lactate to pyruvate in the presence of co-enzyme NAD+. Once cells are impaired by stress, injuries, chemicals, or intercellular signals, LDH is rapidly released from the cell membrane. Thus, the measurement of the amount of released LDH from cells is one of the major methods to assess the cell death. Since Dojindo’s Cytotoxicity LDH Assay Kit-WST neither reflects the activity of living cells nor is harmful to cells, it allows the assay to perform in wells containing both viable and damaged cells.

Choose between Two ProceduresCytotoxicity LDH Assay Kit-WST can be applied with and without supernatant transferring. Please choose suitable method for your experiment.

Assay Procedure[Optimization of cell concentration][Homogeneous Assay][Non-homogeneous Assay]

Stable Working SolutionWorking Solution is stable for 6 months under refrigerated conditions. Therefore, after the preparation, Working Solution can be used as a ready-to-use solution at any time during this period.

Experimental Examples

To obtain an accurate result in cytotoxicity assay, the samples are measured by different principles. Cell Counting Kit-8 (Product Code: CK04) measure NADH in living cells and LDH assay measure released LDH from dead cells. According to these results, the living cells decreased and dead cells increased with higher concentration of toxicant.

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Stability of Working Solution

Reconstituted Working Solution has higher stability than competitor’s kit, eliminating the additional time to prepare the Working Solution for each assay.

Comparison of Working Solution Stability

Linearity

Dojindo’s Cytotoxicity LDH Assay Kit-WST has higher sensitivity than competitor’s kits

Cytotoxicity of mitomycin C using HeLa cells

Culture medium: MEM, 10% FBSIncubation: 37C, 5% CO2, 48 hours

[Optimization of cell concentration][Homogeneous Assay][Non-homogeneous Assay]

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