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Cell Counting Kit 8 (CCK-8)
Cell Counting Kit 8 (CCK-8) is a colorimetric assay used to quantitate cell proliferation, cell viability and cytotoxicity.
Product Description
CCK-8 utilizes the enzymatic activity of live cells to reduce a colorless compound into an orange dye, formazan. The amount of formazan produced, as determined by absorbance (λ460 nm), is directly proportional to the number of live cells in the sample.
Key Features
- Faster results with improved water solubility compared to MTT assays
- Higher sensitivity compared to MTS or WST-1 assays, enabling detection of live cells at low densities
- Lower cytotoxicity compared to XTT based assays, reducing the noise in the data due to assay components
- Excellent stability allowing for long assays (24 - 48 hours)
- Convenient workflow with a single-component, mix-and-read format
- Enhanced accuracy compared to MTT assays, as it targets total dehydrogenase instead of mitochondrial dehydrogenase activity
Assay Principle
Metabolically active cells are known to produce dehydrogenase enzymes. Under neutral pH conditions and with the aid of an electron acceptor known as 1-methoxy phenazine methosulfate, NADPH-dependent cellular enzymes operate across the cell membrane to mediate the reduction of the colorless, cell-impermeant WST-8 tetrazolium salt. This yields an orange-colored formazan dye which is soluble in cell culture media or aqueous buffers, exhibiting strong absorption at 460 nm. This absorbance is directly proportional to the number of live cells present in the samples, serving as a reliable indicator of cell viability.
What is the difference between CCK-8 and MTT?
CCK-8 assays rely on total cell dehydrogenase activity to reduce WST-8 to an orange formazan dye, which is monitored at λ 460 nm.
MTT assays rely on mitochondrial dehydrogenase activity to reduce MTT to a purple formazan dye, which is monitored at λ 562 nm.
CCK-8 assays are less toxic, more sensitive and more water soluble than MTT assays.
MTT assays rely on mitochondrial dehydrogenase activity to reduce MTT to a purple formazan dye, which is monitored at λ 562 nm.
CCK-8 assays are less toxic, more sensitive and more water soluble than MTT assays.
What is the difference between CCK-8 and CellTiter-Glo?
CellTiter-Glo is a luminescence assay that relies on production of oxyluciferin from exogenous luciferin, catalyzed by luciferase in the presence of ATP from metabolically active cells.
CCK-8 is a colorimetric assay that relies on production of formazan from WST-8, catalyzed by endogenous cellular dehydrogenase in the presence of NADPH from metabolically active cells.
CCK-8 is a colorimetric assay that relies on production of formazan from WST-8, catalyzed by endogenous cellular dehydrogenase in the presence of NADPH from metabolically active cells.
What is the formula for CCK-8?
CCK-8 uses WST-8 as the primary reagent for detecting cell viability.
The chemical name of WST-8 is 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H tetrazolium monosodium salt.
The molecular formula for WST-8 is C20H14N6NaO11S2+.
The SMILES of WST-8 is COC1=C(C=CC(=C1)[N+](=O)[O-])[N+]2=NC(=NN2C3=CC=C(C=C3)[N+](=O)[O-])C4=C(C=C(C=C4)S(=O)(=O)O)S(=O)(=O)[O-].[Na+].
The chemical name of WST-8 is 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H tetrazolium monosodium salt.
The molecular formula for WST-8 is C20H14N6NaO11S2+.
The SMILES of WST-8 is COC1=C(C=CC(=C1)[N+](=O)[O-])[N+]2=NC(=NN2C3=CC=C(C=C3)[N+](=O)[O-])C4=C(C=C(C=C4)S(=O)(=O)O)S(=O)(=O)[O-].[Na+].
What is the OD value of CCK-8?
CCK-8 is a colorimetric assay. The assay results can be read using an absorbance spectrophotometer. The resulting optical density (OD) value is proportional to the number of viable cells. The maximum OD value for CCK-8 occurs at 460 nm.
Is CCK-8 light sensitive?
CCK-8 should be stored so as to minimize light exposure. The recommended storage condition is < -15 °C.
References
View all 50 references: Citation Explorer
An investigation on the changes of serum CCK-8, substance P, and 5-HT in patients with post-stroke insomnia.
Authors: Zhang, Xiao-Hua and Zhang, Xin and Feng, Hong-Ye and Cao, Chang-Chun and Lv, Hui-Lan and Wang, Yu-Long and Ren, Li-Jie
Journal: Technology and health care : official journal of the European Society for Engineering and Medicine (2023): 2355-2361
Authors: Zhang, Xiao-Hua and Zhang, Xin and Feng, Hong-Ye and Cao, Chang-Chun and Lv, Hui-Lan and Wang, Yu-Long and Ren, Li-Jie
Journal: Technology and health care : official journal of the European Society for Engineering and Medicine (2023): 2355-2361
Correlation research of serum substance P, CCK-8, and 5-HT values with depression levels in stroke survivors.
Authors: Zhang, X and Wang, C-B and Duan, L-H and Long, J-J and Xiao, P and Wang, Y-L and Zhang, X-H and Liu, Q-Q
Journal: European review for medical and pharmacological sciences (2023): 1248-1254
Authors: Zhang, X and Wang, C-B and Duan, L-H and Long, J-J and Xiao, P and Wang, Y-L and Zhang, X-H and Liu, Q-Q
Journal: European review for medical and pharmacological sciences (2023): 1248-1254
An optimized protocol to detect protein ubiquitination and activation by ubiquitination assay in vivo and CCK-8 assay.
Authors: Lin, Xiao-Tong and Zhang, Jie and Xie, Chuan-Ming
Journal: STAR protocols (2023): 102199
Authors: Lin, Xiao-Tong and Zhang, Jie and Xie, Chuan-Ming
Journal: STAR protocols (2023): 102199
CCK-8 enhances acid-sensing ion channel currents in rat primary sensory neurons.
Authors: Qin, Qing-Rui and Xu, Zhong-Qing and Liu, Ting-Ting and Li, Xue-Mei and Qiu, Chun-Yu and Hu, Wang-Ping
Journal: Neuropharmacology (2023): 109739
Authors: Qin, Qing-Rui and Xu, Zhong-Qing and Liu, Ting-Ting and Li, Xue-Mei and Qiu, Chun-Yu and Hu, Wang-Ping
Journal: Neuropharmacology (2023): 109739
Inhibitory Effect of CCK-8 on Methamphetamine-Induced Apoptosis.
Authors: Zhang, Wu-Hua and Zhang, Ming-Long and Jing, Wei-Wei and Xie, Bing and Bi, Hai-Tao and Yu, Feng and Cong, Bin and Ma, Chun-Ling and Wen, Di
Journal: Fa yi xue za zhi (2021): 796-805
Authors: Zhang, Wu-Hua and Zhang, Ming-Long and Jing, Wei-Wei and Xie, Bing and Bi, Hai-Tao and Yu, Feng and Cong, Bin and Ma, Chun-Ling and Wen, Di
Journal: Fa yi xue za zhi (2021): 796-805
Page updated on September 11, 2024
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