Contents of the Kit: 100 tests
Storage Condition: 0-5°C
|| Assay Buffer
||:11 ml, 1 tube |
||:1.1 ml, 1 tube
|| Stop Solution
||:5.5 ml, 1 tube |
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
Cytotoxicity LDH Assay Kit-WST is a kit for determination of cytotoxicity by measuring a lactate dehydrogenase(LDH) activity released from damaged cells. LDH is a stable cytoplasmic enzyme presented in all types of cells and released into the cell culture medium through damaged plasma membrane. Cytotoxicity LDH Assay kit-WST can be used to measure the released LDH according to the following scheme. LDH catalyzes dehydrogenation of lactate to pyruvate thereby reducing NAD to NADH. NADH reduces a water-soluble tetrazolium salt(WST) in the presence of an electron mediator to produce an orange formazan dye. The amount of the formazan dye thus formed is proportional to that of released LDH into the medium, which is an indication of cytotoxicity.
Since Cytotoxicity LDH Assay kit-WST neither reflects the activity of living cells nor is harmful to cells, cytotoxicity can be measured with the living cells(homogeneous assay). In addition, non-homogeneous assay that is performed by using the cell culture supernatant is also possible. Unlike competitive products, the reconstituted Working Solution is stable under refrigerated condition and the Working Solution can be used for long periods as ready-to-use solution after the preparation.
Fig. 1 LDH detection mechanism
The reconstituted "Working Solution" could be stored for 2 months at refrigerator.
Comparison of Working Solution Stability
Dojindo Cytotoxicity LDH Assay Kit-WST allows longer storage of working solution once it is reconstituted.
Cytotoxicity of mitomycin C using HeLa cells
Culture medium: MEM, 10% FBS
Incubation: 37C, 5% CO2, 48 hours
Stability of Working Solution
Reconstituted Working Solution can be stored for 2 months at 0-5C