CCK-8 - Cell Counting Kit-8 - Cell Proliferation and Cytotoxicity Assay DJDB4000X

Cell Counting Kit-8 (CCK-8) - Cell Proliferation and Cytotoxicity Assay - Cell-Based and Functional Assay Dojindo Molecular Technologies, Inc DJDB4000X
Supplier: Dojindo Molecular Technologies, Inc
Product Code: DJDB4000X
Availability: In Stock

Available Options

* CCK-8 Kit Size:






Cell Counting Kit-8 (CCK-8) allows sensitive colorimetric assays for the determination of cell viability in cell proliferation and cytotoxicity assays. 

  • Simple to use
  • Non-radioactive colorimetric assay 
  • Out-performs old generation assays (e.g. MTT, MTS or XTT)

How it works

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.


Applications:

Non-Radioactive Cell Proliferation or Cytotoxicity Assay

The Cell Counting Kit-8 (CCK8) is a one-bottle kit that requires no premixing.  This non-radioactive, sensitive colorimetric assay allows accurate live cell counting in a cell proliferation or cytotoxicity assay applications.  CCK8 based assay has excellent correlation with results of traditional radioactive [3H]-thymidine incorporation assay.

Other Applicable Assays:

  • Cell viability.
  • Chemosensitivity.
  • Cell attachment determination.
  • Chemotaxis

CCK-8 Out Performs Old Generation Colorimetric Cell Proliferation Compounds: MTT, MTS and XTT

CCK-8 has significantly higher signal


CCK-8 is less toxic to cells than WST-1or MTS:


CCK-8 is easier to use: Just add CCK-8 and measure!


Available Kit Sizes:

CCK-8 Kits are available in the following sizes: 100 tests, 500 tests, 1000 tests, 3000 tests and 10000 tests

 


CCK-8 Documents:

CCK-8 Kit User Manual 

CCK-8 MSDS Sheet 

CCK-8 FAQ 


 

Cell Counting Kit-8 Frequently Asked Questions

  1. 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.

 

  1. 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.

 

  1. 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.

 

  1. 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.

 

  1. 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.

 

  1. 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.

 

  1. 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.

 

  1. How stable is CCK-8?

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

 

  1. 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.


Selected Publications using CCK-8

Tissue Origin  Cell Line  References
bovine brain microvascular endothelial  BBMVEC  T. Kitamuro, et al., J. Biol. Chem., 278, 9125 (2003)
dog canine kidney epithelial cell  MDCK  H. Shimura, et al., Cancer res., 61, 3640 (2001)
human anaplastic thyroid carcinoma  ARO  F. Furuya, et al., Endocrinology, 145, 2865 (2004)
human B lymphoid  WSU-CLL  M. Ho, et al., J. Biol. Chem., 280, 607 (2005)
human bladder carcinoma  T24  Y. Shibata, et al., J. Biol. Chem., 277, 746 (2002)
human bone marrow mesenchymal stem cell  BMMSC  M. Miura, et al., Stem Cells, 24, 1095 (2006)
human bronchial epithelial cell  BEAS-2B  C. A. Reilly, et al., Toxicol. Sci., 73, 170 (2003)
    M. E. Johansen, et al., Toxicol. Sci., 89, 278 (2006)
human burkitt lymphoma  Daudi  M. Ho, et al., J. Biol. Chem., 280, 607 (2005)
  Ramos  M. Ho, et al., J. Biol. Chem., 280, 607 (2005)
human cervical carcinoma  C33A  W. Yang, et al., Mol. Cancer Ther., 5, 1610 (2006)
human colon carcinoma  HCT116  H. Ohori, et al., Mol. Cancer Ther., 5, 2563 (2006)
human colorectal adenocarcinoma  DLD-1  H. Ohori, et al., Mol. Cancer Ther., 5, 2563 (2006)
human embryonal carcinoma  NT2N  J. Tessier, et al., Infect. Immun., 75, 1895 (2007)
human epithelial carcinoma cell  A431  H. Ohori, et al., Mol. Cancer Ther., 5, 2563 (2006)
human esophageal squamous cell carcinoma  KYSE  I. Imoto, et al., Cancer Res., 61, 6629 (2001)
    K. Nakakuki, et al., Carcinogenesis, 23, 19 (2002)
human gastric cancer cell  SH10TC  H. Ohori, et al., Mol. Cancer Ther., 5, 2563 (2006)
human gingival fibroblast  Gin-1  R. Takii, et al., Infect. Immun., 73, 883 (2005)
human glioblastoma  T98G  T. Kitamuro, et al., J. Biol. Chem., 278, 9125 (2003)
  U87MG  S. Kim, et al., Clin. Cancer Res., 11, 5965 (2005)
    S. Kim, et al., Clin. Cancer Res., 12, 5550 (2006)
human intrahepatic bile duct carcinoma cell  HuCCT1  H. Ohori, et al., Mol. Cancer Ther., 5, 2563 (2006)
human kidney carcinoma  293T  H. Fuda, et al., J. Lipid Res., 48, 1343 (2007)
human leukemia cell  Kasumi-1  G. Zhou, et al., Blood, 109, 3441 (2007)
human lung adenocarcinoma  LK87  H. Ohori, et al., Mol. Cancer Ther., 5, 2563 (2006)
human lung cancer cell  H1299  H. Tominaga, et al., Anal. Commun., 36, 47 (1999)
     S. Semba, et al., J. Biol. Chem., 281, 28244 (2006)
human lymphoblast cell  SupT1  J. Melton, et al., J. Biol. Chem., 279, 14315 (2004)
  T-cell  I. Y. Lee, et al., J. Immunol., 175, 1658 (2005)
  Namalwa  M. Ho, et al., J. Biol. Chem., 280, 607 (2005) 
human medulloblastoma  Daoy  X. Li, et al., Mol. Cancer Ther., 4, 1912 (2005)
    S. Kim, et al., Clin. Cancer Res., 12, 5550 (2006)
human mesenchymal stem cell  hMSC  D. Huang, et al., FASEB J., 19, 2014 (2005)
    L. Song, et al., Stem Cells, 24, 1707 (2006)
human monoblastic lymphoma  U937  R. Hori, et al., J. Biol. Chem., 277, 10712 (2002)
human multiple myeloma  AMO1  J. Inoue, et al., Am. J. Pathol., 165, 71 (2004)
  KMS-11  J. Inoue, et al., Am. J. Pathol., 165, 71 (2004)
human neural stem cell  HB1.F3  S. Kim, et al., Clin. Cancer Res., 11, 5965 (2005)
     S. Kim, et al., Clin. Cancer Res., 12, 5550 (2006)
human neuroblastoma  IMR32  H. Tominaga, et al., Anal. Commun., 36, 47 (1999)
  SK-N-SH  Y. Wang, et al., J. Virol., 78, 7916 (2004)
  SMS-KAN  A. Misawa, et al., Cancer Res., 65, 10233 (2005)
human non-small-cell lung cancer cell  LCSC#2  H. Ishibashi, et al., Cancer Res., 65, 6450 (2005)
  RERF-LC-OK  H. Ishibashi, et al., Cancer Res., 65, 6450 (2005)
human ovarial cancer cell  OVK18  H. Ohori, et al., Mol. Cancer Ther., 5, 2563 (2006)
human ovarian adenocarcinoma  HTOA  M. Furuya, et al., Cancer Res., 65, 2617 (2005)
human pancreatic cancer cell  Alexander cell  S. Awale, et al., Cancer Res., 66, 1751 (2006)
  AsPC-1  T. Mori, et al., Mol. Cancer Ther., 3, 29 (2004)
     S. Awale, et al., Cancer Res., 66, 1751 (2006)
  BxPC-3  S. Awale, et al., Cancer Res., 66, 1751 (2006)
  MiaPaCa-2  A. Aghdassi, et al., Cancer Res., 67, 616 (2007)
human peripheral blood mononuclear cell  PBMC  T. Mori, et al., Mol. Cancer Ther., 3, 29 (2004)
    S. Awale, et al., Cancer Res., 66, 1751 (2006)
    A. Aghdassi, et al., Cancer Res., 67, 616 (2007)
     C. Chang, et al., Stem Cells, 24, 2466 (2006)
     T. Lee, et al., Mol. Cancer Ther., 5, 2398 (2006)
human prostate carcinoma  LNCaP  D. J. Son, et al., Mol. Cancer Ther., 6, 675 (2007)
human pulmonary adenocarcinoma  H441  H. Shimura, et al., Cancer res., 61, 3640 (2001)
human skin mast cell  primary Mast cell  J. Tessier, et al., Infect. Immun., 75, 1895 (2007)
human T cell  Jurkat  T. Kasai, et al., J. Biol. Chem., 277, 5187 (2002)
     L. Lu, et al., J. Biochem., 141, 157 (2007)
mouse cortical neurons,  primary  M. Ikonen, et al., PNAS, 100, 13042 (2003)
mouse Macrophage    Y. Miyake, et al., J. Immunol., 178, 5001 (2007)
mouse embryonic fibroblast  Balb3T3  H. Tominaga, et al., Anal. Commun., 36, 47 (1999)
  3T3-L1  D. Huang, et al., FASEB J., 19, 2014 (2005)
mouse fibroblast  L929  H. Tominaga, et al., Anal. Commun., 36, 47 (1999)
  NIH3T3  R. Yu, et al., Toxicol. Sci., 93, 82 (2006)
mouse hippocampal cell  HT22  H. Sohn, et al., FASEB J., 20, 1428 (2006)
mouse insulinoma  MIN6  S. Oyadomari, et al., PNAS, 98, 10845 (2001)
mouse macrophage  RAW 264  M. Shiga, et al., Anesth. Analg., 92, 128 (2001)
  RAW 264.7  S. Oyadomari, et al., PNAS, 98, 10845 (2001)
    D. J. Son, et al., Mol. Cancer Ther., 6, 675 (2007)
mouse hepatocellular carcinoma  MH134  S. Shibata, et al., J. Immunol., 177, 3564 (2006)
mouse malignant melanoma  B16F1  S. Shibata, et al., J. Immunol., 177, 3564 (2006)
Wistar rats calvaria osteoblast     E. Hinoi, et al., FASEB J., 17, 1532 (2003)
General
Size 100 tests, 500 tests, 1000 tests, 3000 tests, 10000 tests
Storage and shelf-life 0-5 °C, keep dry and away from light; 6 month at room temp 1 year at 0-5 °C; >2 year at -20 °C

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