Cytotoxicity LDH Assay Kit-WST


Cytotoxicity Assay Kit-WST
∼ Feature ∼
– Can be used with and without transferring supernatant
– Stable Working Solution (6 months at 0-5 °C)
– No need to freeze the Working Solution

Storage Condition: 0-5°C
Shipping Condition: ambient temperature

Required Equipment and Materials:
Microplate reader with 490 nm filter, CO2 incubator,
96-well tissue culture plate(flat bottomed), 20 μl, 100-200 μl multi-channel pipette

Kit Contents by Unit Size:

DescriptionReferencesDataVideoSupplemental ManualManual for ADCC AssayManualS.D.S

Detection Principle
Lactate 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 Procedures
Cytotoxicity 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 Solution
Working 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.

1) S. F. Jin, H. L. Ma, Z. L. Liu, S. T. Fu, C. P Zhang, Y. He, “XL413, a cell division cycle 7 kinase inhibitor enhanced the anti-fibrotic effect of pirfenidone on TGF-β1-stimulated C3H10T1/2 cells via Smad2/4.”, Exp Cell Res., 2015, 2015113000019.
2) S. Watanabe, C. S. Moniaga, S. Nielsen, M. Hara-Chikuma, “Aquaporin-9 facilitates membrane transport of hydrogen peroxide in mammalian cells.Aquaporin-9 facilitates membrane transport of hydrogen peroxide in mammalian cells.”, Biochem Biophys Res Commun ., 2016, 471, 191.
3) L. Wu, T. Oshima, J. Shan, H. Sei, T. Tomita, Y. Ohda, H. Fukui, J. Watari, H. Miwa , “PAR-2 activation enhances weak acid-induced ATP release through TRPV1 and ASIC sensitization in human esophageal epithelial cells.”, Am J Physiol Gastrointest Liver Physiol ., 2015, 309, G695.
4) T. Fukami, A. Iida, K. Konishi, M. Nakajima , “Human arylacetamide deacetylase hydrolyzes ketoconazole to trigger hepatocellular toxicity”, Biochem. Pharmacol.., 2016, doi:10.1016/j.jphs.2016.07.007.
5) M. Tanaka, M. Yoneyama, T. Shiba, T. Yamaguchi, K. Ogita, “Protease-activated receptor-1 negatively regulates proliferation of neural stem/progenitor cells derived from the hippocampal dentate gyrus of the adult mouse “, J Pharmacol Sci., 2016, doi:10.1016/j.jphs.2016.05.005.
6) S. Wakatsuki, A. Furuno, M. Ohshima, and T. Araki, “Oxidative stress-dependent phosphorylation activates ZNRF1 to induce neuronal/axonal degeneration”, J Cell Biol., 2015, 211, (4), 881.

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


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% FBS
Incubation: 37C, 5% CO2, 48 hours

[Optimization of cell concentration]

[Homogeneous Assay]

[Non-homogeneous Assay]

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