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Cell Counting Kit-8

Item # Unit Size
CK04-11
1,000 tests
CK04-13
3,000 tests
CK04-20
10,000 tests

For Research Use Only Products


Storage Condition : 0-5oC
Shipping Condition : ambient temperature

Product Description
Cell Counting Kit-8 (CCK-8) allows sensitive colorimetric assays for the determination of cell viability in cell proliferation and cytotoxicity assays. Dojindo’s highly water-soluble tetrazolium salt, WST-8, is reduced by dehydrogenase activities in cells to give a yellow-color formazan dye, which is soluble in the tissue culture media. The amount of the formazan dye, generated by the activities of dehydrogenases in cells, is directly proportional to the number of living cells. The detection sensitivity of CCK-8 is higher than the other tetrazolium salts such as MTT, XTT, MTS or WST-1.




Cell viability detection mechanism with CCK-8

Procedure

Cell Counting Kit-8 requires 3 simple steps.

The handling time of Cell Counting Kit-8 is the shortest among its competitors. Only 15 minutes of handling time is needed for Cell Counting Kit-8, whereas longer handling time is required for both MTS and MTT assay.

Unlike MTT assay, there is no need to lyse cells, so you don't have to worry about the data variation.


Sensitivity

CCK-8 (WST-8) is the highest sensitive dye for the cell based assay.
MTT Users, Click Here for differences between CCK-8 and MTT

Comparison of the sensitivity using tetrazolium dye


Cytotoxicity of Reagents

Only in CCK-8, continious culture is possible without killing cells.

Observation of Cytotoxicity after incubation with each reagent.
HeLa cells were incubated with WST-8 (CCK-8), WST-1, or MTS at 24 hours from addition of those reagents.

Stability

Cell Counting Kit-8 is a ready-to-use solution. It is also stable at 4ºC for 1 year. Your assay can be done anytime without thaw and freeze.


Purpose of Using CCK-8 and Cytotoxicity LDH Assay Kit-WST


Because Cell Counting Kit-8 measures the metabolic activity of living cells, the data does not verify cell death. To ensure the data is reflecting the cell death instead of decreased metabolic activity, Cytotoxicity LDH Assay Kit-WST (CK12), which measures LDH released from dead cells, is used to increase the data reliability.

Example Data using Cell Counting Kit-8


Data for Cell Proliferation and Cytotoxicity Assay

Cell Viability Determination



Data was kindly provided by Reaction Biology Corporation.

Absorption property of WST-8



The microplate reader with a 450-490 nm filter is applicable for the measurement.



Absorption spectrum of WST-8 formazan dye

Most Recent
Publications

Cytotoxicity Test for 3D Cultured Model
The cytotoxicity test method for 3D cultured model using Cell Counting Kit-8 has been added to OECD TG492 as the standard method. For details, please Click Here.


Publications based on cell line

Cell Line Description Publication
10T1/2 mouse embryonic fibroblast H. Fung, et al., Mol. Cell. Biol., 27, 8834 (2007)
16HBE-T human bronchial epithelial cell, malignant transformed Y. Jiang, et al., Toxicol. Sci., 119, 50 (2011)
2008/C13*5.25 human ovarian cancer cell J. Song, et al., Clin. Cancer Res., 13, 6842 (2007)
293T human kidney carcinoma H. Fuda, et al., J. Lipid Res., 48, 1343 (2007)
293T human kidney carcinoma B. Zhou, et al., J. Virol., 82, 6962 (2008)
3T3-L1 mouse embryonic fibroblast D. Huang, et al., FASEB J., 19, 2014 (2005)
3T3-L1 mouse embryonic fibroblast T. Shimada, et al., Am. J. Physiol. Endocrinol. Metab., 296, E721 (2009)
3T3-L1 mouse embryonic fibroblast Tsuyoshi Shimada, et al., Am J Physiol Endocrinol Metab, 296, E721 (2009)
3T3-L1 mouse embryonic fibroblast J. Lee, et al., j biol chem, 285, 32647 (2010)
3Y1 rat fibroblastic cell N. Itano, et al., PNAS, 99, 3609 (2002)
786-0 renal carcinoma cell Y. Zhong, et al., Carcinogenesis, 30, 158 (2009)
8505c human anaplastic thyroid carcinoma H. Ohori, et al., Mol. Cancer Ther., 5, 2563 (2006)
A2780 human ovarian cancer cell J. Song, et al., Clin. Cancer Res., 13, 6842 (2007)
A2780cis human ovarian cancer cell J. Song, et al., Clin. Cancer Res., 13, 6842 (2007)
A431 human epithelial carcinoma cell H. Ohori, et al., Mol. Cancer Ther., 5, 2563 (2006)
A431/K5 human epidermoid carcinoma Y. Zhang, et al., Clin. Cancer Res., 12, 4695 (2006)
A431/K5 human epidermoid carcinoma R. Hassan, et al., Clin. Cancer Res., 13, 7166 (2007)
A549 human lung cancer cell T. Shiozaki, et al., J. Gen. Virol., 92, 315 (2011)
A549 human lung cancer cell S. Semba, et al., J. Biol. Chem., 281, 28244 (2006)
A549 human lung cancer cell C. A. Reilly, et al., Toxicol. Sci., 73, 170 (2003)
A549 human lung cancer cell J. Stankova, et al., Clin. Cancer Res., 11, 2047 (2005)
A549 human lung cancer cell H. Ishibashi, et al., Cancer Res., 65, 6450 (2005)
A549 human lung cancer cell I. Imoto, et al., Cancer Res., 66, 4617 (2006)
A549 human lung cancer cell T. Kitamuro, et al., J. Biol. Chem., 278, 9125 (2003)
A549 human lung cancer cell E. Kawata, et al., Mol. Cancer Ther., 7, 2904 (2008)
A549 human lung cancer cell J. Yin, et al., Mol. Pharmacol., 74, 1132 (2008)
A549 human lung cancer cell J. Xu, et al., Carcinogenesis. 31, 927-935 (2010)
A549 human lung cancer cell M. Nguyen, et al., Cancer Res., 70, 5337-5347 (2010)
A549 human lung cancer cell Y.Miki, et al., Cancer Res., 70, 6659-6669(2010)
A549 human lung cancer cell Y. Li, et al., Toxicological Sciences, 116, 1, 174-182(2010)
A6 kidney epithelial cells K. Tamura, et al. Blood. 115. 850-859 (2010)
ACHN human renal adenocarcinoma T. Morikawa, et al., Clin. Cancer Res., 13, 5703 (2007)
ACHN human renal adenocarcinoma H. Ohori, et al., Mol. Cancer Ther., 5, 2563 (2006)
AGS Gastric cancer cell Y. Hayakawa, et al., PNAS, 108, 780 (2011)
AGS Gastric cancer cell Z. Dubeykovskaya, et al., J. Biol. Chem., 284, 3650 (2009)
AGS Gastric cancer cell Q. Yang, et al., Carcinogenesis, 32, 713 (2011)
AGS Gastric cancer cell Siyuan Zhang, et al., Mol Cancer Res, 7, 570 (2009)
Alexander pancreatic cancer cell S. Awale, et al., Cancer Res., 66, 1751 (2006)
ALVA-101 human prostate cancer cell T. Kawakubo, et al., Cancer Res., 67, 10869 (2007)
ALVA-41 human prostate cancer cell T. Kawakubo, et al., Cancer Res., 67, 10869 (2007)
AMO1 multiple myeloma J. Inoue, et al., Am. J. Pathol., 165, 71 (2004)
APL-NB4 acute promyelocytic leukemia cell M. A. S. Khan, et al., PNAS, 101, 11560 (2004)
APRE19 human retinal pigment epithelium E. F. Moreira, et al., Invest. Ophthamol. Vis. Sci., 50, 523 (2009)
AR42J Pancreatic acinar cell R. Szmola, et al., Pancreas. 59, 365-372 (2010)
AR42J Pancreatic acinar cell C. Bose, et al., Am. J. Physiol. Gastrointest. Liver Physiol., 289, G926 (2005)
ARO human anaplastic thyroid carcinoma Z. Meng, et al., Endocrinology, 149, 5357 (2008)
ARO human anaplastic thyroid carcinoma F. Furuya, et al., Endocrinology, 145, 2865 (2004)
ARPE-19 spontaneously arising retinal pigment epithelia cell I. Larrayoz, et al., Inv.Ophthalmology & Visual Science, 51(10), 4942-4955(2010)
ASC adipose-derived stem cell Yuka Ikegame, et al., Cytotherapy, 1-11(2011) online only
AsPC-1 pancreatic cancer cell T. Mori, et al., Mol. Cancer Ther., 3, 29 (2004)
AsPC-1 pancreatic cancer cell S. Awale, et al., Cancer Res., 66, 1751 (2006)
ASTC-a-1 Human Lung Adenocarcinoma cell M. Yu, et al., Experimental Biology and Medicine, 235, 1053-1061 (2010)
ASTC-a-1 Human Lung Adenocarcinoma cell Yingjie Zhang, et al., Molecular Biology of the Cell, 20, 3077 (2009)
ATN-1 T-cell T. Hisatomi, et al., Blood, 117, 3575 (2011)
B16F1 murine malignant melanoma S. Shibata, et al., J. Immunol., 177, 3564 (2006)
B16F10 murine malignant melanoma S. Shibata, et al., J. Immunol., 177, 3564 (2006)
Ba/F3 Lymphoid cell Wenhai Liu, et al., J. Biochem., 145, 575 (2009)
Balb3T3 mouse embryonic cell H. Tominaga, et al., Anal. Commun., 36, 47 (1999)
BBMVEC bovine brain microvascular endothelial T. Kitamuro, et al., J. Biol. Chem., 278, 9125 (2003)
BE(2)-C human neuroblastoma J. Qiao, et al., PNAS, 105, 12891 (2008)
BEAS-2B human bronchial epithelial cell K. C. Thomas, et al., J. Pharmacol. Exp. Ther., 337, 400 (2011)
BEAS-2B human bronchial epithelial cell C. A. Reilly, et al., Toxicol. Sci., 73, 170 (2003)
BEAS-2B human bronchial epithelial cell M. E. Johansen, et al., Toxicol. Sci., 89, 278 (2006)
BEAS-2B human bronchial epithelial cell G. Murphy, Jr., et al., Am. J. Respir. Cell Mol. Biol., 38, 532 (2008)
BEAS-2B human bronchial epithelial cell K. C. Thomas, et al., J. Pharmacol. Exp. Ther., 321, 830 (2007)
BEAS-2B human bronchial epithelial cell ER stress, et al., The J. of pharm. and Exp. Therap., 337, 400-410 (2011)
BEL7404 human hepatoma cell X. Liang, et al., Mol. Cancer Res., 6, 1499 (2008)
BGC-823 human gastric cancer cell Q. Yang, et al., Carcinogenesis, 32, 713 (2011)
BHP15-3 human papillary thyroid carcinoma H. Shimura, et al., Cancer res., 61, 3640 (2001)
BHP18-21v human papillary thyroid carcinoma H. Shimura, et al., Cancer res., 61, 3640 (2001)
BHP18-21v human papillary thyroid carcinoma F. Furuya, et al., Endocrinology, 145, 2865 (2004)
BHP7-13 human papillary thyroid carcinoma H. Shimura, et al., Cancer res., 61, 3640 (2001)
BMM bone marrow macrophage H. J. Kim, et al., Mol. Pharmacol., 72, 418 (2007)
BMMSC bone marrow mesenchymal stem cell M. Miura, et al., Stem Cells, 24, 1095 (2006)
BMSC BM(bone marrow)-derived stem cell Yuka Ikegame, et al., Cytotherapy, 1-11(2011)
bone marrows mouse S. J. Priceman, et al., Blood, 115. 1461 (2010)
BPH-1 human benign prostatic hyperplasia cell F. An, et al., Mol. Cancer Ther., 7, 569 (2008)
brain endothelials mouse D. Zhu, et al., Blood, 115, 4963 (2010)
breast cancer from stage IV breast cancer patient E. I. Chen, et al., Cancer Res., 67, 1472 (2007)
BT-474 human breast cancer cell D. Iliopoulos, et al., Clin. Cancer Res., 13, 268 (2007)
BxPC-3 pancreatic cancer cell S. Awale, et al., Cancer Res., 66, 1751 (2006)
C2C12 mouse myoblast cell J. Sakamaki, et al., PNAS, 108, 6085 (2011)
C2C12 mouse myoblast cell W. Yoon, et al., J. Biol. Chem., 283, 32751 (2008)
C3 Rat mammary cancer cell J. Xu, et al., Carcinogenesis. 31, 927-935 (2010)
C33A human cervical carcinoma W. Yang, et al., Mol. Cancer Ther., 5, 1610 (2006)
C6 rat glioblastoma cell M. Zhao, et al., Anticancer Res., 30, 2217-2224 (2010)
C6 rat glioblastoma cell Wen Jing Lu, et al., Int. J. Cancer, 128, 343-351(2011)
C8161.9 human melanoma cell A. A. Dar, et al., J. Biol. Chem., 286, 16606 (2011)
C8166 T-cell T. Kasai, et al., J. Biol. Chem., 277, 5187 (2002)
C8166-45 T-cell J. Peloponese, Jr., et al., J. Virol., 83, 3238 (2009)
CA46 human burkitt lymphoma J. E. Weldon, et al., Blood, 113, 3792 (2009)
Ca9-22 human gingival epithelial cell N. Ebe, et al., J. Dent. Res., 90, 235 (2011)
Caco-2 human caucasian colon adenocarcinoma T. Suzuki, et al., J. Nutr., 141, 87 (2011)
Caco-2 cel Human intestinal cell Takuya Suzuki, et al., J. Nutr., 139, 965 (2009)
Caki-1 renal carcinoma T. Morikawa, et al., Clin. Cancer Res., 13, 5703 (2007)
Caki-2 renal carcinoma T. Morikawa, et al., Clin. Cancer Res., 13, 5703 (2007)
Caov-3 human serous ovarian cancer cell H. Yoshida, et al., Anticancer Res., 31, 1271 (2011)
cardiomyocyte rat H. Y. Fu, et al., Cardiovasc. Res., 79, 600 (2008)
cardiomyocyte rat E. E. Hamm, et al., PNAS, 103, 14176 (2006)
cardiomyocyte rat K. Shintani-Ishida, et al., Am. J. Physiol. Heart Circ. Physiol., 293, H1714 (2007)
CDC cardiac-derived stem cell J. Terrovitis, et al., J. AM. Coll. Cardiol. 54, 1619-1626 (2010)
CDC cardiac-derived stem cell J. Terrovitis, et al., Circulation, 117, 1555 (2008)
cerebellar granule neuron rat X. Wang, et al., J. Biol. Chem., 280, 16705 (2005)
CGC rat goblet cell J. D. Rios, et al., Invest. Ophthalmol. Vis. Sci., 48, 1543 (2007)
CHO chinese hamster ovary cell A. Kunita, et al., Am. J. Pathol., 170, 1337 (2007)
CHO chinese hamster ovary cell S. Yokoe, et al., Cancer Res., 67, 1935 (2007)
chondrocyte from human cartilage tissue T. Sato, et al., Ann. Rheum. Dis., 70, 221 (2011)
chondrocyte, primary from mouse embryo Y. Shinoda, et al., J. Biol. Chem., 283, 24682 (2008)
CLL chronic lymphocytic leukemia John E. Weldon, et al., Blood, 113, 3792 (2009)
CLL chronic lymphocytic leukemia V. El-Khooury, et al., Mol. Cancer. Ther. 9, 1349-1360 (2010)
CMECs mouse myocardial microvascular endothelial cell Jiming Li, et al., Int.J.Mol.Sci., 12, 1306-1315(2011)
CNE2 and T-47Ds Tumor cell Yu Wang, et al., Cancer Res, 69, 5194 (2009)
COLO 205 Human Caucasian colon adenocarcinoma Y. V. Kovtun, et al., Cancer Res, 70, 2528 (2010)
Colo320DM human colon cancer cell M. Koyama, et al., Mol. Cancer Ther., 6, 1579 (2007)
COR-L23/P human large-cell lung tumor cell C. Wu, et al., Mol. Cancer Ther., 6, 3287 (2007)
cortical neuron mouse Y. Wang, et al., Stroke, 40, 1864 (2009)
COS-7 African green monkey kidney cell H. Bando, et al., Clin. Cancer Res., 11, 5784 (2005)
COS-7 African green monkey kidney cell T. Kitamuro, et al., J. Biol. Chem., 278, 9125 (2003)
COS-7 African green monkey kidney cell N. Sato, et al., Clin. Cancer. Res. 16, 226-239 (2010)
COS-7 African green monkey kidney cell Tsuyoshi Kimura, et al., Journal of Drug Delivery, DOI 10.1155/2011/962743
CV-1 African green monkey kidney cell R. Fukuda, et al., J. Biol. Chem., 284, 2680 (2009)
CV-1 African green monkey kidney cell J. Peloponese, et al., J. Biol. Chem., 281, 8927 (2006)
D283-MED human medulloblastoma X. Li, et al., Mol. Cancer Ther., 4, 1912 (2005)
Daoy human medulloblastoma S. Kim, et al., Clin. Cancer Res., 12, 5550 (2006)
Daoy human medulloblastoma X. Li, et al., Mol. Cancer Ther., 4, 1912 (2005)
Daudi human burkitt lymphoma J. E. Weldon, et al., Blood, 113, 3792 (2009)
Daudi human burkitt lymphoma M. Ho, et al., J. Biol. Chem., 280, 607 (2005)
Daudi human burkitt lymphoma M. Onda, et al., PNAS, 105, 11311 (2008)
DDLS dedifferentiated liposarcoma G. Ambrosini, et al., Mol. Cancer Ther., 7, 890 (2008)
Dermal stem HR-1 mice Yuichi Hasebe, et al., Journal of Dermatological Science, 62, 98-106(2011)
DLD-1 human colorectal adenocarcinoma M. Koyama, et al., Clin Cancer Res, 16, 2320 (2010)
DLD-1 human colorectal adenocarcinoma H. Ohori, et al., Mol. Cancer Ther., 5, 2563 (2006)
DLD-1 human colorectal adenocarcinoma M. Koyama, et al., Clin. Cancer. Res. 16, 2320-2332 (2010)
DLD-1 human colorectal adenocarcinoma S. Kamiyama, et al., Glycobiology, 21, 235 (2011)
DT40 chicken B-lymphocyte cell F. Shinozaki, et al., J. Biol. Chem., 281, 16361 (2006)
DU145 human prostate carcinoma T. Kawakubo, et al., Cancer Res., 67, 10869 (2007)
DU145 human prostate carcinoma D. J. Son, et al., Mol. Cancer Ther., 6, 675 (2007)
DU145 human prostate carcinoma P. Davis-Searles, et al., Cancer Res., 65, 4448 (2005)
E14TG2a mouse embryonic stem cell N. Sasaki, et al., J. Biol. Chem., 283, 3594 (2008)
EC human embryonal carcinoma X. Wang, et al., Stem Cells, 27, 568 (2009)
EC9706 esophageal squamous cell carcinoma Z. Chen, et al., J. Biol. Chem., 286, 10725 (2011)
ESC endometriotic stromal cell T. Hirata, et al., Endocrinology, 149, 1260 (2008)
ES-E14TG2a Murine ES cell Yeong-Min Yoo, et al., Brain Research, 1385, 1-7(2011)
F11 Neurohybrid cells Yuichi Hashimoto, et al., Molecular Biology of the Cell, 20, 2864 (2009)
F98 Brainstem glioma, Human NSC S. Choi, et al., Neuro-Oncology, 13, 61-69(2011)
F98s Brainstem glioma Do-Hun Lee, et al., Clin Cancer Res, 15, 4925 (2009)
FRCC001 renal carcinoma cell Y. Zhong, et al., Carcinogenesis, 30, 158 (2009)
FRCC562 renal carcinoma cell Y. Zhong, et al., Carcinogenesis, 30, 158 (2009)
FRH0201 portal cholangiocellular carcinoma cell N. Guo, et al., Acta Biochem Biophys Sin, 43, 354 (2011)
FRO human anaplastic thyroid carcinoma Z. Meng, et al., Endocrinology, 149, 5357 (2008)
FRT rat thyroid cell H. Shimura, et al., Cancer res., 61, 3640 (2001)
FRTL-5 rat thyroid cell H. Shimura, et al., Cancer res., 61, 3640 (2001)
G361 human melanoma cell H. Ohori, et al., Mol. Cancer Ther., 5, 2563 (2006)
GCIY human gastric carcinoma H. Ohori, et al., Mol. Cancer Ther., 5, 2563 (2006)
GES-1 human gastric epithelial cell Q. Yang, et al., Carcinogenesis, 32, 713 (2011)
Gin-1 human gingival fibroblast R. Takii, et al., Infect. Immun., 73, 883 (2005)
GSM06 gastric surface mucous cell T. Nakajima, et al., Am. J. Physiol. Gastrointest. Liver Physiol., 282, G359 (2002)
H1299 human lung cancer cell E. Kawata, et al., Mol. Cancer Ther., 7, 2904 (2008)
H1299 human lung cancer cell S. Semba, et al., J. Biol. Chem., 281, 28244 (2006)
H1299 human lung cancer cell H. Yamada, et al., Carcinogenesis, 28, 2013 (2007)
H1299 human lung cancer cell Y. Su, et al., Cancer Res., 67, 3329 (2007)
H1299 human lung cancer cell H. Tominaga, et al., Anal. Commun., 36, 47 (1999)
H1650 human non-small-cell lung cancer cell J. Deng, et al., Cancer Res., 67, 11867 (2007)
H1703 non-small-cell lung cancer cell Y. Su, et al., Cancer Res., 67, 3329 (2007)
H1975 human non-small-cell lung cancer cell T. Okabe, et al., Clin. Cancer Res., 15, 907 (2009)
H1975 human non-small-cell lung cancer cell J. Deng, et al., Cancer Res., 67, 11867 (2007)
H23 human lung cancer cell Heekyoung Yang, et al., Biochem. and Biophys. Res. Com., 406, 53-58(2011)
H3255 human non-small-cell lung cancer cell J. Deng, et al., Cancer Res., 67, 11867 (2007)
H441 human pulmonary adenocarcinoma H. Shimura, et al., Cancer res., 61, 3640 (2001)
H9c2 embryonic rat heart-derived cell T. Okazaki, et al., Am. J. Pathol., 171, 1093 (2007)
H9c2 embryonic rat heart-derived cell M. Kimura, et al., J. Biol. Chem., 276, 26453 (2001)
HaCaT human keratinocyte cell H. Lee, et al., J. Immunol., 186, 1248 (2011)
HaCaT human keratinocyte cell H. Yue, et al., J. Immunol. 183, 6839-6848 (2010)
HaCaT human keratinocyte cell Y. Li, et al., Toxicological Sciences, 116, 1, 174-182(2010)
HAK-1A human hepatocellular carcinoma S. Oie, et al., Mol. Cancer Ther., 6, 2310 (2007)
hAMSCs human amniotic mesenchymal stem cell J. Lee, et al., j biol chem, 285, 32647 (2010)
hASM human airway smooth muscle cell C. J. Delvecchio, et al., Am. J. Physiol. Lung Cell Mol. Physiol., 295, L949 (2008)
HB1.F3 human neural stem cell S. Kim, et al., Clin. Cancer Res., 12, 5550 (2006)
HB1.F3 human neural stem cell S. Kim, et al., Clin. Cancer Res., 11, 5965 (2005)
HCC Hepatocellular carcinoma Q. Deng, et al., Cancer Res. 70, 782-791 (2010)
HCC Hepatocellular carcinoma Pan Yue, et al., European Journal of Cancer, 47, 1115-1124(2011)
HCC Hepatocellular carcinoma M. Furuta, et al., Carcinogenesis. 31, 766-776 (2010)
HCC1937 human breast cancer cell D. Iliopoulos, et al., Clin. Cancer Res., 13, 268 (2007)
HCC827 human non-small-cell lung cancer cell J. Deng, et al., Cancer Res., 67, 11867 (2007)
HCC827 GR5 human non-small-cell lung cancer cell T. Okabe, et al., Clin. Cancer Res., 15, 907 (2009)
HCC827 GR6 human non-small-cell lung cancer cell T. Okabe, et al., Clin. Cancer Res., 15, 907 (2009)
HCECs Human Corneal Epithelial Cell N. C. Joyce, et al., Invest Ophthalmol Vis Sci, 50, 2116 (2009)
HCL lymphocytic leukemia, John E. Weldon, et al., Blood, 113, 3792 (2009)
HCT116 human colon cancer cell Jung Ok Ban, et al., Mol Cancer Ther, 8, 1613-1624(2009)
HCT116 human colon cancer cell H. Ohori, et al., Mol. Cancer Ther., 5, 2563 (2006)
HCT116 human colon cancer cell J. Lu, et al., Cancer Res., 68, 9384 (2008)
HCT116 human colon cancer cell M. Iwaizumi, et al., Gut, 58, 249 (2009)
HCT116 human colon cancer cell A. N, Tse, et al., Mol. Pharmacol., 75, 124 (2009)
HCT116 human colon cancer cell M. Tanaka, et al., Anticancer Res., 31, 763 (2011)
HCT116 human colon cancer cell H. Fung, et al., J. Biol. Chem., 286, 4968 (2011)
HCT116 human colon cancer cell Z. Huang, et al., Cancer Res., 71, 2582 (2011)
HCT116 human colon cancer cell Y. Zhao, et al., Pnas. 107, 2592-2597 (2010)
HCT116 human colon cancer cell C. A. Glazer, et al., Arch. Otolaryngol. Head Neck Surg., 137, 286 (2011)
HCT-8 human colon carcinoma Z. Huang, et al., Cancer Res., 71, 2582 (2011)
HEK293 human embryonic kidney cell J. Hyung, et al., Mol. Cancer Res. 8, 246-253 (2010)
HEK293 human embryonic kidney cell S. Semba, et al., J. Biol. Chem., 281, 28244 (2006)
HEK293 human embryonic kidney cell C. Wu, et al., Mol. Cancer Ther., 6, 3287 (2007)
HEK293 human embryonic kidney cell C. Tanikawa, et al., Mol Cancer Res, 8, 855-863(2010)
HEK293/4.63 human embryonic kidney cell C. Wu, et al., Mol. Cancer Ther., 6, 3287 (2007)
HEK293/5I human embryonic kidney cell C. Wu, et al., Mol. Cancer Ther., 6, 3287 (2007)
HEK293T human embryonic kidney cell J. Sakamaki, et al., PNAS, 108, 6085 (2011)
HeLa human cervical carcinoma Y. Seo, et al., Cancer Res. 69, 8356-8365 (2010)
HeLa human cervical carcinoma S. Ramakrishna, et al., J. Biol. Chem., 286, 10505 (2011)
HeLa human cervical carcinoma C. Shi, et al., J. Biol. Chem., 279, 17224 (2004)
HeLa human cervical carcinoma H. Shimura, et al., Cancer res., 61, 3640 (2001)
HeLa human cervical carcinoma H. Tominaga, et al., Anal. Commun., 36, 47 (1999)
HeLa human cervical carcinoma M. Ishiyama, et al., Talanta, 44, 1299 (1997)
HeLa human cervical carcinoma Suresh Ramakrishna, et al., J.of Bio. Chem., 286, 12, 10505-10514(2011)
HeLa human cervical carcinoma R.Minami, et al., J. Cell Biol., 190(4), 637-650 (2010)
HeLa human cervical carcinoma H. Ariyama, et al., Journal of Biological Chem., 285(29) 22027-22035 (2010)
Hep3B human hepatocellular carcinoma J. K. Sicklick, et al., Carcinogenesis, 27, 748 (2006)
hepatocyte Main tissue of liver Y. Jung, et al., Hepatology. 59, 655-665 (2010)
HepG2 Human hepatocellular liver carcinoma cell F. Luan, et al., Hepatology. 58, 1528-1537 (2010)
HepG2 Human hepatocellular liver carcinoma cell Xin-Rong Yang, et al., Clin Cancer Res, 15, 5518-5527(2009)
HepG2 Human hepatocellular liver carcinoma cell H. Ohori, et al., Mol. Cancer Ther., 5, 2563 (2006)
HepG2 Human hepatocellular liver carcinoma cell C. A. Reilly, et al., Toxicol. Sci., 73, 170 (2003)
HepG2 Human hepatocellular liver carcinoma cell T. Shiokawa, et al., Clin. Cancer Res., 11, 2018 (2005)
HepG2 Human hepatocellular liver carcinoma cell T. Ohuchida, et al., Cancer, 100, 2430 (2004)
HepG2 Human hepatocellular liver carcinoma cell H. E. Rasmussen, et al., J. Nutr., 138, 476 (2008)
HepG2 Human hepatocellular liver carcinoma cell L.Liu, et al., ABBS, 42(8)585-592 (2010)
HepG2 Human hepatocellular liver carcinoma cell Ayumi Oshikata, et al., J. of Bio. and Bioeng., 111, 5, 590-593(2011)
HEY human ovarian cancer cell J. Song, et al., Clin. Cancer Res., 13, 6842 (2007)
HEYC2 human ovarian cancer cell J. Song, et al., Clin. Cancer Res., 13, 6842 (2007)
hGal9 modified human Galectin-9 J. Kurida, et al., Mol Cancer Res, 8, 994-1001 (2010)
Hippocampal CA1 neuronal cell Yuichi Kanmura, et al., Anesth Analg. 100, 888-894 (2010)
HL60 human acute promyelonic leukemia A. Kanayama, et al., J. Leukoc. Biol., 82, 1344 (2007)
HL60 human acute promyelonic leukemia H. Tominaga, et al., Anal. Commun., 36, 47 (1999)
HL60 human acute promyelonic leukemia Jie Xiao, et al., Acta Biochim Biophys Sin, 41, 708 (2009)
hMSC human mesenchymal stem cell L. Song, et al., Stem Cells, 24, 1707 (2006)
hMSC human mesenchymal stem cell D. Huang, et al., FASEB J., 19, 2014 (2005)
hMSC human mesenchymal stem cell Ji-Soo Kim, et al., Experimental Gerontology, 46, 500-510(2011)
hMVEC human dermal microvascular endothelial R. Yu, et al., Toxicol. Sci., 93, 82 (2006)
hMVEC human dermal microvascular endothelial K. Kuwabara, et al., Endocrinology, 145, 3324 (2004)
hMVEC human dermal microvascular endothelial E. F. Moreira, et al., Invest. Ophthamol. Vis. Sci., 50, 523 (2009)
hMVEC human dermal microvascular endothelial F. Lahargue, et al., Blood, 115, 5418-5426 (2010)
HO-1-N-1 oral squamous cell carcinoma K. Kozaki, et al., Cancer Res., 68, 2094 (2008)
hPASMC pulmonary artery smooth muscle cells R. Zhang, et al., Am. J. Physiol Lung Cell Mol. Physiol. 297, L631-L640 (2009)
HSC-2 oral squamous cell carcinoma K. Kozaki, et al., Cancer Res., 68, 2094 (2008)
HSC-6 oral squamous cell carcinoma K. Kozaki, et al., Cancer Res., 68, 2094 (2008)
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MIN6 mouse insulinoma S. Oyadomari, et al., PNAS, 98, 10845 (2001)
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mononuclear from bone marrow J. Choi, et al., Blood, 109, 471 (2007)
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NHBE normal human bronchial epithelial cell ER stress, et al., The Journal of pharmacology and Experimental Therapeutics, 337, 400-410, 2011
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NHF normal human fibroblast E. Kawata, et al., Mol. Cancer Ther., 7, 2904 (2008)
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SK-N-BE human neuroblastoma T. Nakagawa, et al., Mol. Cell. Biol., 22, 2575 (2002)
SK-NFI human neuroblastoma J. Stankova, et al., Clin. Cancer Res., 11, 2047 (2005)
SK-N-SH human neuroblastoma Y. Wang, et al., J. Virol., 78, 7916 (2004)
SK-N-SH, SH-SY5Y-Luc, IMR-32, LAN-1, JF, SH-EP, SMS-KCN, SK-N-AS, MCF-10A, MCF-7 human neuroblastoma, Breast normal epithelial, Breast tumor Xiaoying Shang, et al., Mol Cancer Ther, 8, 2461 (2009)
SM43 mesangial cell S. Endo, et al., Mol. Pharmacol., 72, 1337 (2007)
SMC-PV pulmonary vascular smooth muscle cell W. Zhou, et al., Am. J. Physiol. Heart Circ. Physiol., 292, H2773 (2007)
SMS-KAN human neuroblastoma A. Misawa, et al., Cancer Res., 65, 10233 (2005)
SNU638 human gastric cancer cell In-Sung song,et al.,Cancer Res, 69, 3131-3139(2009)
SPC mouse splenocytes C. Zhang, et al., J. Biol. Chem., 286, 13723 (2011)
splenic lymphocytes broiler chickens A. Y. Han, et al., Poultry Science, 89, 2063 (2010)
splenocyte from mouse T. Wu, et al., J. Virol., 81, 10310 (2007)
ST1 T-cell T. Hisatomi, et al., Blood, 117, 3575 (2011)
ST-8814 malignant peripheral nerve sheath tumor cell G. Ambrosini, et al., Mol. Cancer Ther., 7, 890 (2008)
SUIT-2 pancreatic cancer cell T. Mori, et al., Mol. Cancer Ther., 3, 29 (2004)
SupT1 human lymphoblast cell J. Melton, et al., J. Biol. Chem., 279, 14315 (2004)
SW1990, BxPC3 Human pancreatic cancer cell line Peng Wang, et al., Carcinogenesis, 30,1497 (2009)
SW480 human colorectal carcinoma Z. Huang, et al., Cancer Res., 71, 2582 (2011)
SW620 human colon cancer cell S. H. Lee, et al., Mol. Cancer Ther., 6, 2786 (2007)
SW620 human colon cancer cell Jung Ok Ban, et al., Mol Cancer Ther, 8, 1613-1624(2009)
SW620 human colon cancer cell H. Ohori, et al., Mol. Cancer Ther., 5, 2563 (2006)
SW620 human colon cancer cell J. Stankova, et al., Clin. Cancer Res., 11, 2047 (2005)
SW839 renal carcinoma T. Morikawa, et al., Clin. Cancer Res., 13, 5703 (2007)
T24 human bladder carcinoma Y. Shibata, et al., J. Biol. Chem., 277, 746 (2002)
T24 human bladder carcinoma M. Miyake, et al., J. Pharmacol. Exp. Ther. 332, 795-802 (2010)
T-47D Breast carcinoma cell K. Takagi, et al., Endocr-relat. Cancer. 17, 415-430 (2010)
T98G human glioblastoma T. Kitamuro, et al., J. Biol. Chem., 278, 9125 (2003)
Tca8113 human tongue squamous cell carcinoma T. Yu, et al., Mol. Cancer Res., 9, 161 (2011)
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Questions:
How many cells should there be in a well?
Can this kit be used with a 384-well plate?
Can this kit be used with a 24-well plate?
Does CCK-8 stain viable cells?
Does phenol red affect the assay?
Is there a correlation between CCK-8 and the Thymidine incorporation assay?
Is CCK-8 toxic to cells?
How stable is CCK-8?
I do not have a 450 nm filter. What other filters can I use?


Answers:
How many cells should there be in a well?
For adhesive cells, at least 1,000 cells are necessary per well (100 μl medium) when using a standard 96-well plate.
For leukocytes, at least 2,500 cells are necessary per well (100 μl medium) because of low sensitivity.
The recommended maximum number of cells per well for the 96-well plate is 25,000. If 24-well or 6-well plates are used for this assay, please calculate the number of cells per well accordingly, and adjust the volume of the CCK-8 solution in each well to 10% of the total volume.

Can this kit be used with a 384-well plate?
Yes, you can use this kit for a 384-well plate. Add CCK-8 solution with 1/10 volume of the culture medium in a well. If the volume of CCK-8 to be added is too small, dilute CCK-8 in half with the medium and add 1/5 of the volume of culture medium in a well.

Can this kit be used with a 24-well plate?
Yes. Add CCK-8 solution with 1/10 volume of the culture medium in a well.


Does CCK-8 stain viable cells?

No, it does not stain viable cells because a water-soluble tetrazolium salt (WST-8) is used for the CCK-8 solution. The electron mediator, 1-Methoxy PMS, receives electrons from viable cells and transfers the electrons to WST-8 in the culture medium. Since its formazan dye is also highly water soluble, CCK-8 cannot be utilized for cell staining.



Does phenol red affect the assay?

No. The absorption value of phenol red in culture medium can be removed by subtracting the absorption value of a blank solution from the absorption value of each well. Therefore, culture medium containing phenol red can be used for the CCK-8 assay.



Is there a correlation between CCK-8 and the Thymidine incorporation assay?

Yes. However, please note that since CCK-8 uses a different assay mechanism from that of the Thymidine assay, so the results may differ. Comparison data are shown in the technical manual, which is available at www.dojindo.com/tm.


Is CCK-8 toxic to cells?

Since the toxicity of CCK-8 is so low, the same cells can be used for other cell proliferation assays such as the crystal violet assay, neutral red assay or DNA fluorometric assay after the CCK-8 assay is completed.



How stable is CCK-8?

CCK-8 is stable over 6 months at room temperature and one year at 0-5 oC with protection from light. For longer storage, we recommend storing the solution at -20 oC ; CCK-8 is stable over 2 years at -20 oC.



I do not have a 450 nm filter. What other filters can I use?

You can use filters with an absorbance between 450 nm and 490 nm.