Cell counting kit 8 (cck8)

Manufactured by Dojindo Laboratories

Sourced in Japan, United States, China, Germany, United Kingdom

The Cell Counting Kit-8 is a colorimetric assay for the determination of cell viability and cytotoxicity. It utilizes a water-soluble tetrazolium salt that produces a water-soluble formazan dye upon reduction in the presence of an electron carrier. The amount of the formazan dye generated is directly proportional to the number of living cells.

Automatically generated - may contain errors

Lab products found in correlation

Microplate reader, by Thermo Fisher Scientific (182 mentions) Microplate reader, by Bio-Rad (176 mentions) Microplate reader, by Agilent Technologies (103 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (95 mentions) Multiskan fc, by Thermo Fisher Scientific (78 mentions) 96 well plate, by Corning (56 mentions) Dimethyl sulfoxide (dmso), by Merck Group (38 mentions) Microplate reader, by Molecular Devices (37 mentions) Microplate reader, by Tecan (34 mentions) Multiskan go, by Thermo Fisher Scientific (34 mentions) Spectramax m5, by Molecular Devices (33 mentions) Elx800, by Agilent Technologies (33 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (30 mentions) Imark microplate reader, by Bio-Rad (29 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (28 mentions) Cck 8, by Dojindo Laboratories (27 mentions) Model 680, by Bio-Rad (24 mentions) Matrigel, by BD (23 mentions) Lipofectamine rnaimax, by Thermo Fisher Scientific (22 mentions) Prism 5, by GraphPad (22 mentions)

5 369 protocols using cell counting kit 8 (cck8)

Neuroprotective Effects of TubA and FGF-21

Check if the same lab product or an alternative is used in the 5 most similar protocols

Starting at seven days in vitro (DIV-7), cortical neurons were pre-treated with 0.1, 0.25, 0.5, 0.75 and 1 μM TubA for two days, and 10 μM glutamate was added at DIV-9. Cell viability was assessed using a Cell Counting Kit-8 (Dojindo Molecular Technologies, Rockville, MD) 24 hours after glutamate treatment. Cell viability results were expressed as percentage of the vehicle-treated control.
Starting at DIV-2, cortical neurons were pre-treated with 1, 2, 5, 7 and 10 nM recombinant human FGF-21 protein (PeproTech, Rocky Hill, NJ) for seven days, and 10 μM glutamate was added at DIV-9. Cell viability was assayed 24 hours later using a Cell Counting Kit-8 (Dojindo Molecular Technologies).
To test whether there is a synergistic effect by combined pre-treatment with TubA and FGF-21, starting at DIV-2, cortical neurons were pre-treated with 1, 2, or 5 nM recombinant human FGF-21 protein for five days, and at DIV-7, 0.25 or 0.5 μM of TubA was added. At DIV-9, 10 μM glutamate was added for 24 hours. Cell viability was evaluated by using a Cell Counting Kit-8 (Dojindo Molecular Technologies).

Wang Z., Leng Y., Wang J., Liao H.M., Bergman J., Leeds P., Kozikowski A, & Chuang D.M. (2016). Tubastatin A, an HDAC6 inhibitor, alleviates stroke-induced brain infarction and functional deficits: potential roles of α-tubulin acetylation and FGF-21 up-regulation. Scientific Reports, 6, 19626.

+ Open protocol

+ Expand

Culturing Skin Cells for Research

Check if the same lab product or an alternative is used in the 5 most similar protocols

NHEKs obtained from neonatal foreskin were purchased from Lonza (Basel, Switzerland) and cultured in keratinocyte growth medium (KBM Gold) with BulletKit (Lonza, Walkersville, MD, USA) containing insulin, human epidermal growth factor, bovine pituitary extract, hydrocortisone, epinephrine, transferrin, and gentamicin/amphotericin B. The cells were serially passaged until 70–80% confluence was achieved, which was no more than three times. Cell proliferation and cytotoxicity were measured with a Cell Counting Kit-8 (Dojindo, Kumamoto, Japan). Moderately pigmented human epidermal melanocytes were purchased from Cascade Biologics (Portland, OR, USA) and maintained and passaged in Medium 254 (#M254500) supplemented with Human Melanocyte Growth Supplement (Life Technologies, Carlsbad, CA, USA), 10% fetal bovine serum, 100 U/mL of penicillin G, and 100 μg/mL of streptomycin sulfate. The human epidermal melanocytes were incubated at 37 °C with 5% CO₂ and regularly passaged at a density of 80% (1:8 ratio). Cell viability was measured using the Cell Counting Kit-8 (Dojindo, Kumamoto, Japan).

Kim J., Lee J, & Choi H. (2021). Intense Pulsed Light Attenuates UV-Induced Hyperimmune Response and Pigmentation in Human Skin Cells. International Journal of Molecular Sciences, 22(6), 3173.

+ Open protocol

+ Expand

Cell Proliferation Assays: EdU Incorporation and CCK-8

Check if the same lab product or an alternative is used in the 5 most similar protocols

MDA-MB-231 cells grown on 96-well plate were cultured in triplicate in 96-well plates at a density of 1×10³ for 12 h. Then, cells were labeled with 30 µM 5-ethynyl-2′-deoxyuridine (Ribobio, Guangzhou, China) for 1 h at 37°C. The cells were fixed with 50 µl 4% formaldehyde for 30 min and 0.5% Triton X-100 for 20 min for permeabilization. One hundred microliters 1× Apollo reaction cocktail was added to each well for 30 min after washing with PBS thrice. Then, cells were stained with 100 µl Hoechst 33342 for 30 min and washed with PBS. The stained cells were visualized under a fluorescent microscope (Olympus Corporation, Tokyo, Japan) and counted with Photoshop (Adobe Systems, Inc., San Jose, CA, USA). The incorporation rate represents EdU fluorescence against DNA content.
(2 (link)). Cell Counting Kit-8 (Dojindo Laboratories, Kumamoto, Japan) was employed to assess cell proliferation. In brief, cells were seeded on a 96-well cell culture cluster (Corning Inc., Corning, NY, USA) at a concentration of 2×10⁴/well in a volume of 100 µl, and grown overnight. Cell Counting Kit-8 (Dojindo) reagents were then added, incubated for 2 h at 37°C, and absorbance was quantified on an automated plate reader. Each experiment was performed in triplicate and repeated at least three times.

Sheng C., Qiu J., Wang Y., He Z., Wang H., Wang Q., Huang Y., Zhu L., Shi F., Chen Y., Xiong S., Xu Z, & Ni Q. (2018). Knockdown of Ran GTPase expression inhibits the proliferation and migration of breast cancer cells. Molecular Medicine Reports, 18(1), 157-168.

+ Open protocol

+ Expand

Effects of AGEs and HQ on RPE Cell Viability

Check if the same lab product or an alternative is used in the 5 most similar protocols

ARPE-19 and h1RPE7 cells (0.5–2.0×10⁴ cells/100 µL in 96-well plate) were incubated with the addition of AGEs and/or HQ for 24 h. After the treatment, the viable cell numbers were determined by a Cell Counting kit-8 (Dojindo Laboratories, Mashikimachi, Japan) according to the manufacture's instructions as described [20] (link), [21] (link), [22] (link). Briefly, WST-8 (2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazoliummonosodium salt) solution was added to cells in 96-well plates, and the cells were incubated at 37 °C for 1–2 h. The optical density of each well was read at 450 nm (reference wave length at 650 nm) using a Sunrise^TM microplate reader (Tecan, Männedorf, Switzerland).
To investigate effects of VEGF-A on cell growth in RPE cells, ARPE-19 cells (2.0×10⁴ cells/100 µL in 96-well plate) were incubated with AGEs, HQ, in the presence of VEGF-A/VEGF-A receptor inhibitors (10 µg/mL sulochrin (Sigma-Aldrich, St. Louis, MO) [23] (link), 3 nM Ki8751 (Calbiochem^®) [24] (link) or 50 nM CBO-P11 (Calbiochem^®) [25] (link)), or siRNA against VEGF or receptor for AGE (RAGE) for 24 h. After the treatment, the viable cell numbers were determined by a Cell Counting kit-8 (Dojindo) according to the manufacture's instructions.

Tsujinaka H., Itaya-Hironaka A., Yamauchi A., Sakuramoto-Tsuchida S., Ota H., Takeda M., Fujimura T., Takasawa S, & Ogata N. (2015). Human retinal pigment epithelial cell proliferation by the combined stimulation of hydroquinone and advanced glycation end-products via up-regulation of VEGF gene. Biochemistry and Biophysics Reports, 2, 123-131.

+ Open protocol

+ Expand

Cell Proliferation Assay with CCK-8

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cell proliferation was measured using Cell Counting Kit-8 (Dojindo, Kumamoto, Japan).In accordance with the manufacturer's instructions for Cell Counting Kit-8, harvested cells were seeded in 96-well plates at 1×10³ per well (n = 5 for each time point) in a final volume of 100 μL. Cells were cultured for 24, 48, 72, and 96 hours after transfection. CCK-8 solution (10 μL) was added into each well, and the absorbance at 450 nm was measured after incubation for 2 hours at 37°C to calculate the number of viable cells.

Yu L., Ding G.F., He C., Sun L., Jiang Y, & Zhu L. (2014). MicroRNA-424 Is Down-Regulated in Hepatocellular Carcinoma and Suppresses Cell Migration and Invasion through c-Myb. PLoS ONE, 9(3), e91661.

+ Open protocol

+ Expand

Evaluating the Anticancer Potential of FK-3000 in Breast Cancer Cell Lines

Check if the same lab product or an alternative is used in the 5 most similar protocols

The human breast carcinoma cell lines MDA-MB-231 and MCF-7 were obtained from the Korean Cell Line Bank (Seoul, Korea). Cells were cultivated in RPMI-1640 (Gibco/BRL, Grand Island, NY, USA) containing 10% fetal bovine serum (Gibco/BRL), 2 mg/ml sodium bicarbonate (Gibco/BRL), 100 U/ml penicillin (Gibco/BRL), and 100 μg/ml streptomycin (Gibco/BRL).
Cells were seeded in 96-well plates (1.5×10⁴ cells/well) and incubated at 37°C in a 5% CO₂ atmosphere. To determine the IC₅₀ of FK-3000, MDA-MB-231 and MCF-7 cells were treated with 0.1% dimethyl sulfoxide (DMSO; vehicle only control) (Sigma-Aldrich Co.) or FK-3000 (0–5 μg/ml) 24 h after seeding. Cell proliferation was analyzed after 24 and 48 h using the cell counting kit-8 (Dojindo Molecular Technologies, Rockville, MD, USA) according to the manufacturer’s instructions.
To evaluate cell viability, cells were treated with 0.1% DMSO, the 48 h IC₅₀ of FK-3000 (0.52 μg/ml for MDA-MB-231 cells; 0.77 μg/ml for MCF-7 cells), 5.0 μM trans-1-(4-hydroxy-cyclohexyl)-4-(4-fluorophenyl)-5-(2-methoxypyridimidin-4-yl)-imidazole (p38 MAPK inhibitor SB 239063, Sigma-Aldrich Co.), or cotreated with the 48 h IC₅₀ of FK-3000 and 5.0 μM SB 239063. Cell viability was evaluated after 48 h using the cell counting kit-8 (Dojindo Molecular Technologies). All experiments were performed in quadruplicate on different days.

LI Y.C., KIM B.H., CHO S.C., BANG M.A., KIM S, & PARK D.H. (2014). 6,7-di-O-acetylsinococuline (FK-3000) induces G2/M phase arrest in breast carcinomas through p38 MAPK phosphorylation and CDC25B dephosphorylation. International Journal of Oncology, 46(2), 578-586.

+ Open protocol

+ Expand

Quantifying Cell Viability with WST-8

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

The WST-8 assay [using Cell Counting Kit-8 (Dojindo Laboratories, Kumamoto, Japan)] is a modified MTT assay that measures the mitochondrial reduction capacity and can quantify cell viability.^{(29 (link))} After treatment with daidzein or 5-hydroperoxyeicosatetraenoic acid (5-HpETE), the cells in 96-well plates were incubated with 10 µl of Cell Counting Kit-8 solution {containing with 4-[3-(2-methoxy-4-nitrophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]-1,3-benzene disulfonate sodium salt} in medium for 1 h at 37°C. Absorbance was measured photometrically at 450 nm. Cell viabilities were expressed as a percentage of the absorbance measured in non-treated cells.

Horio Y., Sogabe R., Shichiri M., Ishida N., Morimoto R., Ohshima A, & Isegawa Y. (2020). Induction of a 5-lipoxygenase product by daidzein is involved in the regulation of influenza virus replication. Journal of Clinical Biochemistry and Nutrition, 66(1), 36-42.

+ Open protocol

+ Expand

Cell Proliferation Assessment Using CCK-8

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cell proliferation ability was assessed by Cell Counting Kit-8 (Dojindo, Japan) at 0 h, 24 h, 48 h and 72 h following the instructions of manufacture. Cells were washed by PBS, and then 10 μL CCK-8 and 90 μL serum free medium were added. After incubation for 2 h under the condition of 95% air plus 5% CO2 at 37 °C, cell proliferation rate was assessed according to the optical density (OD) value (450 nm) detected by Microplate reader (BioTek, USA). Cell Counting Kit-8 (Dojindo, Japan).

Ren L., Chen S., Liu W., Hou P., Sun W, & Yan H. (2019). Downregulation of long non-coding RNA nuclear enriched abundant transcript 1 promotes cell proliferation and inhibits cell apoptosis by targeting miR-193a in myocardial ischemia/reperfusion injury. BMC Cardiovascular Disorders, 19, 192.

+ Open protocol

+ Expand

Quantifying A549 Cell Viability via WST-8 Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

The viability of A549 cells was quantified by the Total Superoxide Dismutase Assay Kit with WST-8 (WST-8 assay) (Tahara et al., 2017 (link)) using a Cell Counting Kit-8 (Dojindo). Cells were incubated in 10% of WST-8 reagent in a volume of 100 μL at 37°C and 5% v/v CO₂ for 30min. The absorbance was measured at 450 nm by a multimode microplate reader (PerkinElmer).
The cytotoxicity of Asprellcosides B was assessed by the WST-8 assay using a Cell Counting Kit-8 (Dojindo). A549 cells were grown in 96-well plates at 37°C and 5% v/v CO₂ for 24 h and then incubated with different concentrations of Asprellcosides B for 48 h. The absorbance was then measured at 450 nm using a multimode microplate reader.

Zhang W., Chen S.T., He Q.Y., Huang L.Q., Li X., Lai X.P., Zhan S.F., Huang H.T., Liu X.H., Wu J, & Li G. (2019). Asprellcosides B of Ilex asprella Inhibits Influenza A Virus Infection by Blocking the Hemagglutinin- Mediated Membrane Fusion. Frontiers in Microbiology, 9, 3325.

+ Open protocol

+ Expand

Cell Proliferation, Adhesion, Invasion, and Wound Healing

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cell proliferation was measured using Cell Counting Kit-8 (Dojindo Laboratories, Kumamoto, Japan) following manufacturer’s instructions. For the cell adhesion assay, 96-well plates were coated with fibronectin (10 μg/ml) at 4°C for 18 h and cells were allowed to adhere for 1.5 hours at 37°C. At the end of this time period, adherent cells were quantified using the Cell Counting Kit-8 (Dojindo Laboratories, Kumamoto, Japan) following the manufacturer’s instructions. Cell invasion and wound healing assays were performed as previously described.^{6 (link)}

Han T.S., Hur K., Xu G., Choi B., Okugawa Y., Toiyama Y., Oshima H., Oshima M., Lee H.J., Kim V.N., Chang A.N., Goel A, & Yang H.K. (2014). MicroRNA-29c-mediates initiation of gastric carcinogenesis by directly targeting ITGB1. Gut, 64(2), 203-214.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!