01 Cell Proliferation / Cell Cytotoxicity Assay

Cytotoxicity LDH Assay Kit-WST

Cytotoxicity Assay

Can be used with and without transferring supernatant
Stable Working Solution (6 months at 0-5 oc)
No need to freeze the Working Solution

Product code

CK12　 Cytotoxicity LDH Assay Kit-WST

Unit size	Price	Item Code
100 tests		CK12-01
500 tests		CK12-05
2000 tests		CK12-20

Component

100 tests	・Dye Mixture ・Assay Buffer ・Lysis Buffer ・Stop Solution	×1 11 ml×1 1.1 ml×1 5.5 ml×1
500 tests	・Dye Mixture ・Assay Buffer ・Lysis Buffer ・Stop Solution	×1 55 ml×1 5.5 ml×1 27.5 ml×1
2000 tests	・Dye Mixture ・Assay Buffer ・Lysis Buffer ・Stop Solution	×4 55 ml×4 5.5 ml×4 27.5 ml×4

C of A

SDS download

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.

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.

Developer	Dojindo Molecular Technologies, Inc.

Manual

Homogeneous Assay

Non-homogeneous Assay

Optimization of cell concentration

Technical info

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

References

Open References

Reference No.	Sample	Citation
1)	Cell （C3H10T1/2）	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, 339, (2), 289.
2)	Cell （HepG2）	S. Watanabe, C. S. Moniaga, S. Nielsen, M. Hara-Chikuma, "Aquaporin-9 facilitates membrane transport of hydrogen peroxide in mammalian cells.", Biochem Biophys Res Commun ., 2016, 471, 191.
3)	Cell （HEECs, Human Esophageal Epithelial Cells）	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)	Cell （Human L02 Hepatocyte)	D. Zhou, B-H Li, J. Wang, Y-N Ding, Y. Dong, Y-W Chen and J-G Fan, "Prolyl Oligopeptidase Inhibition Attenuates Steatosis in the L02 Human Liver Cell Line.", PloS ONE., 2016, 11, (10), e0165224.
5)	Cell （Human primary Hepatocyte)	T. Fukami, A. Iida, K. Konishi, M. Nakajima , "Human arylacetamide deacetylase hydrolyzes ketoconazole to trigger hepatocellular toxicity", Biochem. Pharmacol., 2016, 116, 153.
6)	Cell （Mouse embryonic fibroblast:MEF）	Y. Takanezawa, R. Nakamura, Y. Sone, S. Uraguchi and M. Kiyono, "Atg5-dependent autophagy plays a protective role against methylmercury-induced cytotoxicity", Toxicol. Lett., 2016, 262, 135.
7)	Cell （neural stem/progenitor cells）	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, 131, (3), 162
8)	Cell （Primary cultured cortical neurons）	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.
9)	Cell （DLD1 and AGS cells）	A. Ishijima, K. Minamihata, S. Yamaguchi, S. Yamahira, R. Ichikawa, E. Kobayashi, M. Iijima, Y. Shibasaki, T. Azuma, T. Nagamune & I. Sakuma, "Selective intracellular vaporisation of antibody-conjugated phase-change nano-droplets in vitro", Scientific Reports ., 2017, DOI:10.1038/srep44077.
10)	Cell （U87, HUVEC cell)	N. Li, W. Zhang, M. Khan, L. Lin, JM. Lin, "MoS2-LA-PEI nanocomposite carrier for real-time imaging of ATP metabolism in glioma stem cells co-cultured with endothelial cells on a microfluidic system", Biosens Bioelectron., 2017, 99, (2018), 142.
11)	Tissue （Mouse liver）	Y. Sakurai, W. Mizumura, M. Murata, T. Hada, S. Yamamoto, K. Ito, K. Iwasaki, T. Katoh, Y. Goto, A. Takagi, M. Kohara, H. Suga, and H. Harashima, "Efficient siRNA delivery by lipid nanoparticles modified with a non-standard macrocyclic peptide for EpCAM-targeting", Mol. Pharm.., 2017,10.1021/acs.molpharmaceut.7b00362.
12)	Cell （Human pleural mesothelial cell)	K. Hattori, K. Nakadate, A. Morii, T. Noguchi, Y. Ogasawara, K. Ishii., "Exposure to nano-size titanium dioxide causes oxidative damages in human mesothelial cells: The crystal form rather than size of particle contributes to cytotoxicity.", Biochem. Biophys. Res. Commun.., 2017,10.1016/j.bbrc.2017.08.054.
13)	Tissue （murine distal colon)	H. Sakai, S. Tabata, M. Kimura, S. Yabe, Y. Isa, Y. Kai, F. Sato, T. Yumoto, K. Miyano, M. Narita and Y. Uezono, "Active Ingredients of Hange-shashin-to, Baicalelin and 6-Gingerol,　Inhibit 5-Fluorouracil-Induced Upregulation of CXCL1 in the Colon to　Attenuate Diarrhea Development", Biol. Pharm.Bull.., 2017, 40, (12), 2134.
14)	Cell （SH-SY5Y cell)	R. Watanabe, T. Kurose, Y. Morishige and K. Fujimori, "Protective Effects of Fisetin Against 6-OHDA-Induced Apoptosis by Activation of PI3K-Akt Signaling in Human Neuroblastoma SH-SY5Y Cells.", Neurochem. Res.., 2018, 43, (2), 488-499.
15)	Cell (MC3T3-E1[Derived from mouse cranial crown])	A. Oyane, M. Nakamura, I. Sakamaki, Y. Shimizu, S. Miyata and H. Miyaji, "Laser-assisted wet coating of calcium phosphate for surface-functionalization of PEEK", PLoS ONE ., 2018, doi:10.1371/journal.pone.0206524.
16)	Cell CRC Cell [Colorectal cancer]、HT29 Cell[Adenocarcinoma of the human colon]	T. Fuji, Y. Umeda, A. Nyuya, F. Taniguchi, T. Kawai, K. Yasui, T. Toshima, K. Yoshida, T. Fujiwara, A. Goel and T.Nagasaka, "Detection of Circulating MicroRNAs with Ago2 Complexes to Monitor the Tumor Dynamics of Colorectal Cancer Patients during Chemotherapy.", Int. J. Cancer., 2018, doi: 10.1002/ijc.31960 .
17)	CEll (SFXN2 knockout HEK293)	E. E. Mon, F. Y. Wei, R. N. R. Ahmad, T. Yamamoto, T. Moroishi and K. Tomizawa, "Regulation of mitochondrial iron homeostasis by siderofexin 2 ", J Physiol Sci., 2018, doi:10.1007/s12576-018-0652-2.
18)	Cell (Human umbilical vein cell line [EA.Hy926])	Y. Xu, C. Huang, M. Liu, N. Chen, W. Chen, C. Yang, Y. Zhao, X. Li, J. Duan, S. Liu and S. Yang, "Survivin regulated by autophagy mediates hyperglycemia-induced vascular endothelial cell dysfunction", Exp. Cell Res.., 2018, doi: 10.1016/j.yexcr.2018.01.037.
19)	Cell (MIAPaCa-2[Human Pancreatic Cancer])	M. Akimoto, R. Maruyama, Y. Kawabata, Y. Tajima and K. Takenaga, "Antidiabetic adiponectin receptor agonist AdipoRon suppresses tumour growth of pancreatic cancer by inducing RIPK1/ERKdependent necroptosis", Cell Death Dis., 2018, 9, 804.
20)	Cell (Fibro adipogenic progenitor cells:FAPs)	Y. Saito, T. Chikenji, T. Matsumura, M. Nakano and M. Fujimiya, "Exercise enhances skeletal muscle regeneration by promoting senescence in fibro-adipogenic progenitors", Nat. Commun., 2020, doi:10.1038/s41467-020-14734-x.

Handling and storage condition

Handling and storage condition
0-5°C, Protect from light
Danger / harmful symbol mark

SDS download

Cytotoxicity LDH Assay Kit-WST

Cytotoxicity LDH Assay Kit-WST