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Cell counting kit 8 cck 8 assay kit

Manufactured by Dojindo Laboratories
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Sourced in Japan, United States, China

The Cell Counting Kit-8 (CCK-8) assay kit is a colorimetric assay for the determination of cell viability and cytotoxicity. The kit uses a water-soluble tetrazolium salt that is reduced by dehydrogenase activities in living cells, producing a colored formazan dye that can be measured spectrophotometrically.

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Lab products found in correlation

Microplate reader, by Agilent Technologies (4 mentions) Microplate reader, by Bio-Rad (4 mentions) Dimethyl sulfoxide (dmso), by Merck Group (4 mentions) Infinite m200, by Tecan (3 mentions) Elisa reader, by Agilent Technologies (3 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions) Victor3 1420 multilabel counter, by PerkinElmer (2 mentions) Elx800, by Agilent Technologies (2 mentions) Cell counting kit 8 cck 8, by Dojindo Laboratories (2 mentions) Multiskan spectrum, by Thermo Fisher Scientific (2 mentions) Bca protein assay kit, by Beyotime (2 mentions) Calcein am, by Merck Group (2 mentions) Microplate computer software, by Bio-Rad (2 mentions) Normocin, by InvivoGen (2 mentions) 96 well microplate, by Corning (2 mentions) Prism 8, by GraphPad (2 mentions) Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions) Sybr green pcr master mix, by Thermo Fisher Scientific (2 mentions) Multiskan go, by Thermo Fisher Scientific (2 mentions)

Spelling variants of this product

CCK-8 (3556 citations) CCK-8 kit (1386 citations) CCK-8 assay (1174 citations) Cell Counting Kit (156 citations) Counting kit (CCK-8; (2 citations)

124 protocols using cell counting kit 8 cck 8 assay kit

1

Evaluating Cell Viability with CCK-8 Assay

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The cell viability was examined using Cell Counting Kit-8 (CCK-8) assay kits (Dojindo Molecular Technologies, Shanghai, China) according to the manufacturer’s instructions. Cells were seeded in 96-cell plates at 1 × 104 cells per well in 100 μL of standard culture medium and incubated overnight. After cells were treated with H2O2 at indicated concentrations, 10 μL of CCK-8 reagent was added to each well and incubated in a tissue culture incubator at 37 °C for 1 h. The absorbance at 450 nm was determined, using a Bio-Tek800 microplate reader (BioTek Instruments, Winooski, VT, USA). Three wells of cells were used for each condition for every independent experiment. The cell viability in H2O2-treated cells was presented as a percentage of that in control or untreated cells in parallel experiments.
An X., Fu Z., Mai C., Wang W., Wei L., Li D., Li C, & Jiang L.H. (2019). Increasing the TRPM2 Channel Expression in Human Neuroblastoma SH-SY5Y Cells Augments the Susceptibility to ROS-Induced Cell Death. Cells, 8(1), 28.
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2

Evaluating Cell Viability with CCK-8 Assay

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Cell viability was determined using Cell Counting Kit-8 (CCK-8) assay kits (Dojindo Molecular Technologies, Inc., Rockville, MD) according to the manufacturer’s instructions. THP-1 cells were seeded at 20,000 cells per well in 96-well plates and differentiated in 200 nM PMA for 72 hr and allowed to rest for 48 hr prior to treatment. [36 ,37 ] Cells were exposed to fresh-prepared cdPM (33 μg/mL) and incubated for 24 hr. The media containing particles were removed, and the cells were incubated in 100 μL of a medium solution containing 10% (v/v) CCK-8 reagent. The cells were incubated at 37°C for a further 3 hr. The absorbance was obtained by scanning with a SpectraMax M2 microplate reader (Molecular Devices, Sunnyvale, CA) at 450 nm. Each experiment was performed at least three times in triplicate. The cells treated with 10% DMSO served as a positive control group, and cells treated with the 15% (volume based) DI water in media served as the negative control. Cell viability was calculated according to the following formula: cell proliferation (%) = absorbance of the experimental group/absorbance of the negative control group ×100. The results were confirmed by visual inspection.
Kaur K., Jaramillo I.C., Mohammadpour R., Sturrock A., Ghandehari H., Reilly C., Paine R I.I.I, & Kelly K.E. (2019). Effect of collection methods on combustion particle physicochemical properties and their biological response in a human macrophage-like cell line. Journal of environmental science and health. Part A, Toxic/hazardous substances & environmental engineering, 54(12), 1170-1185.
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3

Cell Viability Assay for HCC Cells

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HCC cell viability was assessed using Cell Counting Kit-8 (CCK-8) assay kits (Dojindo, Nagasaki, Japan) as described previously5 (link). In brief, 4,000 cells were seeded into 96-well culture plates and allowed to attach for 6 h. Then, after 0, 24, 48, and 72 h, 100 mL of CCK-8 assay buffer was added to each well. The optical density (450 nm) was measured in each well using a microplate reader (Bio Tek, Winooski, VT, USA) according to the manufacturer’s instructions. Three independent experiments were performed.
He M., Hu J., Fang T., Tang W., Lv B., Yang B, & Xia J. (2022). Protein convertase subtilisin/Kexin type 9 inhibits hepatocellular carcinoma growth by interacting with GSTP1 and suppressing the JNK signaling pathway. Cancer Biology & Medicine, 19(1), 90-103.
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4

Gambogic Acid-Induced Apoptosis Mechanisms

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GA (≥98% purity; molecular formula, C30H16O4; molecular weight, 470.69; CAS No. 471-53-4) was purchased from Chengdu Pufei De Biotech Co., Ltd., China. GA was dried under reduced pressure at 60°C for 2 hours before use, dissolved in DMSO, and used immediately after preparation. Cell Counting Kit-8 (CCK-8) assay kits were obtained from Dojindo (Japan). The Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) apoptosis detection kits were obtained from BestBio Institute of Biotechnology (Shanghai, China). Antibodies against Keap1 (AF5266, 1 : 1000 dissolution), Nrf2 (AF0639, 1 : 1000 dissolution), HO-1 (AF5393, 1 : 1000 dissolution), Bax (AF0120, 1 : 2000 dissolution), Bcl-2 (AF6139, 1 : 1000 dissolution), PCNA (AF0239, 1 : 5000 dissolution), β-actin (TA0022, 1 : 3000 dissolution), and GAPDH (AF7021, 1 : 5000 dissolution) were obtained from Affinity Biosciences (OH, USA). Cell Signaling Technology, Inc. (Boston, MA, USA) provided the primary antibodies against cleaved Caspase-3 (9664, 1 : 1000 dissolution) and PARP (9532s, 1 : 1000 dissolution). Abcam (Cambridge, UK) provided the antibodies against caspase-9 (ab184186, 1 : 2000 dissolution).
Jiang Z., Wang Y., Xi X., Cai W., Liu C., Ye R., Yang L., Zhang S., Zhang R., Xu Q, & Yang L. (2022). Regulation of the Keap1-Nrf2 Signaling Axis by Glycyrrhetinic Acid Promoted Oxidative Stress-Induced H9C2 Cell Apoptosis. Evidence-based Complementary and Alternative Medicine : eCAM, 2022, 2875558.
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5

Cell Proliferation Assay with CCK-8

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Cell proliferation was measured using Cell Counting Kit-8 (CCK-8) assay kits (Dojindo Corp., Kumamoto, Japan) according to the manufacturer’s instructions. Huh7 cells and MHCC-97H cells (5×103) were placed in 96-well plates (1,000 cells per well) and incubated at 37°C for 24 h.
Li K., Niu Y., Li K., Zhong C., Qiu Z., Yuan Y., Shi Y., Lin Z., Huang Z., Zuo D., Yuan Y, & Li B. (2023). Dysregulation of PLOD2 Promotes Tumor Metastasis and Invasion in Hepatocellular Carcinoma. Journal of Clinical and Translational Hepatology, 11(5), 1094-1105.
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6

Cell Viability Assay Using CCK-8

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Cell viability was assessed using the Cell Counting Kit-8 (CCK-8) assay kit from Dojindo Laboratories (Kumamoto, Japan). In brief, stably transfected and control cells were seeded into a 96-well culture plate at a density of 1×103 cells/well and cultured for up to five days, with media replaced every three days. At the end of each experiment, 5 µl of the CCK-8 reagent was added into each well and the cells were further cultured for 4 h. Then, the optical density of cells was measured at 450 nm using a spectrophotometer (Tecan, Männedorf, Switzerland). The experiments were performed in triplicate and repeated at least three times.
Chen Y., Zhao G., Li N., Luo Z., Wang X, & Gu J. (2019). Role of 4-aminobutyrate aminotransferase (ABAT) and the lncRNA co-expression network in the development of myelodysplastic syndrome. Oncology Reports, 42(2), 509-520.
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7

Cell Viability Assessment with CCK8 Assay

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A Cell Counting Kit-8 (CCK8) assay kit (Dojindo, Kumamoto, Japan) was used as previously described [12 (link)]. Briefly, cells were plated in 96-well plates at a density of 4 × 103 cells/well. Then, cells were cultured for 48 h, and 10 μl of CCK8 solution was added to the cells and incubated for 2.5 h at 37°C. The absorbance was measured at 450 nm using a microplate reader (Infinite M200, TECAN, Switzerland).
Teng S., Ma C., Yu Y, & Yi C. (2019). Hydroxyurea promotes TET1 expression and induces apoptosis in osteosarcoma cells. Bioscience Reports, 39(5), BSR20190456.
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8

Cell Viability and Oxidative Stress

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Cell viability and LDH were determined using Cell Counting Kit-8 (CCK-8) assay kit (catalogue no. 04-11; Dojindo, Kumamoto, Japan) and cytotoxicity assay kit (catalogue no. A020-2-2; Jiancheng, Nanjing, China), according to the manufacturer's protocols. Cells SOD and 15-F2t-isoprostane releases which reflect oxidative stress level were detected using assay kits according to the manufacturer's protocols.
Bin Z., Yanli Y., Zhen Q., Qingtao M, & Zhongyuan X. (2020). GDF11 ameliorated myocardial ischemia reperfusion injury by antioxidant stress and up-regulating autophagy in STZ-induced type 1 diabetic rats. Acta Cirúrgica Brasileira, 34(11), e201901106.
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9

Cell Proliferation Determination by CCK8

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For cell proliferation assays, cells were seeded into each well of a 96-well plate (2000 per well) and the cell proliferation ability was determined by the Cell Counting Kit-8 (CCK8) Assay Kit (Dojindo Corp, Japan). 10ul of the kit reagent dissolved with 100μl DMEM was added to each well, and 2h later the absorbance was measured at 450 nm to calculate the number of cells.
Yu T., Liu L., Li J., Yan M., Lin H., Liu Y., Chu D., Tu H., Gu A, & Yao M. (2015). MiRNA-10a is upregulated in NSCLC and may promote cancer by targeting PTEN. Oncotarget, 6(30), 30239-30250.
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10

Bupivacaine-Induced Oxidative Stress in SH-SY5Y Cells

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The human dopaminergic neuroblastoma SH-SY5Y cell line was purchased from the Shanghai Institutes for Biological Sciences. Bupivacaine hydrochloride (purity 99.9%), glucose (purity 99.5%), and N-acetyl-L-cysteine (NAC) were purchased from Sigma (St. Louis, MO). Other reagents used included Dulbecco's modified Eagle medium (DMEM)/F12 (including 17.5 mM glucose) and fetal bovine serum (FBS: Gibco, Grand Island, NY); Cell Counting Kit-8 (CCK8) assay kit (Dojindo, Kumamoto, Japan); 2′,7′-dichlorofluorescein diacetate (DCFH-DA) (Beyotime, China); 4′,6-diamidino-2-phenylindole dihydrochloride n-hydrate (DAPI) which were purchased from Wako Pure Chemical Industries Ltd. (Osaka, Japan); comet assay (Trevigen, Inc., Gaithersburg, MD); anti-OGG1 and anti-8-oxodG (Abcam, Cambridge, UK, ab115841 and ab62623); anti-caspase-9 (CST, USA, 9508) and anti-β-actin (KangChen Bio-tech, China, KC-5A08). All reagents were obtained from commercial suppliers and were of standard biochemical quality.
Liu Z.J., Zhao W., Zhang Q.G., Li L., Lai L.Y., Jiang S, & Xu S.Y. (2015). OGG1 Involvement in High Glucose-Mediated Enhancement of Bupivacaine-Induced Oxidative DNA Damage in SH-SY5Y Cells. Oxidative Medicine and Cellular Longevity, 2015, 683197.
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