Cell viability was measured using CCK-8 cell proliferation kit (Dojindo). Ten µL CCK-8 reagent was added into 100 µL cell culture medium. After incubation for 2.5 hours in 37 °C, the absorbance values were measured at 450 nm with the microplate reader (Molecular Devices).
Cck 8 cell proliferation kit
Manufactured by Dojindo Laboratories
Sourced in Japan
The CCK-8 cell proliferation kit is a colorimetric assay for the determination of cell viability and proliferation. It utilizes the highly water-soluble tetrazolium salt WST-8, which is reduced by dehydrogenases in viable cells to produce a yellow-colored formazan dye. 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
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Microplate reader, by Molecular Devices (2 mentions)
Annexin v 7 aad apoptosis detection kit, by Keygen Biotech (2 mentions)
Microplate reader, by Bio-Rad (2 mentions)
Cisplatin, by Bio-Techne (1 mentions)
Beckman cytoflex, by Beckman Coulter (1 mentions)
Herceptin, by Roche (1 mentions)
Infinite 200 pro, by Tecan (1 mentions)
Cytoflex flow cytometer, by Beckman Coulter (1 mentions)
Anisomycin, by Merck Group (1 mentions)
Facscalibur flow cytometry, by BD (1 mentions)
Recombinant human interleukin il 6, by R&D Systems (1 mentions)
Stattic sc 202818, by Santa Cruz Biotechnology (1 mentions)
Ultrapure water, by Merck Group (1 mentions)
Trypsin, by Thermo Fisher Scientific (1 mentions)
Edu cell proliferation kit with alexa fluor 488, by Beyotime (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions)
Fluorescence microscope, by Zeiss (1 mentions)
5 ethynyl 2 deoxyuridine edu assay kit, by RiboBio (1 mentions)
50 mm edu, by RiboBio (1 mentions)
21 protocols using cck 8 cell proliferation kit
1
CCK-8 Cell Viability Assay
2
Evaluating SH3BGRL's Role in Anticancer Drug-Induced Apoptosis
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Cells were seeded in 96-well plates at a density of 1000 cells per well. The growth rate of cells was evaluated by using the CCK-8 cell proliferation kit (Dojindo Laboratories, Kumamoto, Japan), according to the manufacturers’ instructions. Cell-cycle analysis was carried out by flow cytometry (Beckman CytoFLEX, California, USA) after propidium iodide staining.
For analysis the role of SH3BGRL in anticancer drug-induced apoptosis, parental MCF-7 and SH3BGRL-overexpressing MCF-7 cells were treated with 10 μM Cisplatin (TOCRIS, Abingdon, OX, UK) for 12 h or with 150 ng Herceptin (Roche, Basel, Switzerland) for 96 h, respectively. Likely, MDA-MB-453 and MDA-MB-453 SH3BGRL knockdown cells were treated with 10 μM Cisplatin for 12 h, with 150 ng Herceptin for 96 h, PI3K/AKT inhibitor LY294002 (CST, Danvers, MA,USA) for 24 h, or Herceptin and ATK inhibitor combination for 48 h, respectively. Cell apoptosis was analyzed using the Annexin V/7-AAD Apoptosis Detection kit (Keygen Biotech, Nanjing, China), and the percentage of apoptotic cells was analyzed by flow cytometer (Beckman CytoFLEX, California, USA). The detailed procedures of all above assays were described as previously [16 (link)].
For analysis the role of SH3BGRL in anticancer drug-induced apoptosis, parental MCF-7 and SH3BGRL-overexpressing MCF-7 cells were treated with 10 μM Cisplatin (TOCRIS, Abingdon, OX, UK) for 12 h or with 150 ng Herceptin (Roche, Basel, Switzerland) for 96 h, respectively. Likely, MDA-MB-453 and MDA-MB-453 SH3BGRL knockdown cells were treated with 10 μM Cisplatin for 12 h, with 150 ng Herceptin for 96 h, PI3K/AKT inhibitor LY294002 (CST, Danvers, MA,USA) for 24 h, or Herceptin and ATK inhibitor combination for 48 h, respectively. Cell apoptosis was analyzed using the Annexin V/7-AAD Apoptosis Detection kit (Keygen Biotech, Nanjing, China), and the percentage of apoptotic cells was analyzed by flow cytometer (Beckman CytoFLEX, California, USA). The detailed procedures of all above assays were described as previously [16 (link)].
3
Colorimetric Cell Viability Assay
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To measure the cell viability, CCK-8 cell proliferation kit (Dojindo, Tokyo, Japan) was used. Cells were diluted to a concentration of 10
3 cells per 100 μL, distributed to the 96-well plate (6 wells, 100 μL per well) and placed in an incubator at 37°C with 5% CO
2 overnight. From day 1 to day 6, 10 μL of CCK-8 reagent was added into each well and placed in an incubator at 37°C with 5% CO
2 for 1 h before measuring the absorbance at 450 nm.
3 cells per 100 μL, distributed to the 96-well plate (6 wells, 100 μL per well) and placed in an incubator at 37°C with 5% CO
2 overnight. From day 1 to day 6, 10 μL of CCK-8 reagent was added into each well and placed in an incubator at 37°C with 5% CO
2 for 1 h before measuring the absorbance at 450 nm.
4
Cell Proliferation and Apoptosis Evaluation
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Cells were seeded in 96-well plates at a density of 2,000 cells per well. The growth rate of cells was evaluated using the CCK-8 cell proliferation kit (Dojindo Laboratories, Kumamoto, Japan), according to the manufacturers’ instructions. OD detection at 450 nm was carried out by infinite 200Pro (Tecan). Cell apoptosis was analyzed using the Annexin V/7-AAD Apoptosis Detection kit (Keygen Biotech, Nanjing, China) on a CytoFLEX flow cytometer (Beckman, California, United States). Results were further analyzed by Flowjo 10.4.
5
BDNF-Releasing Hydrogel Enhances PC12 Proliferation
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PC12 cells were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). Cells were cultured in DMEM (Gibco®; Thermo Fisher Scientific, Waltham, MA, USA) with 10% fetal bovine serum (Thermo Fisher Scientific), 0.1 mg/mL streptomycin, and 100 U/mL penicillin. Cultures were incubated at 37°C, 5% CO2, and 95% humidity. The cultivated medium was replaced every 2 days. Cells were passaged when confluence reached 80%–90%. HAMC-KAFAK/BDNF treatment was used to evaluate the effect of released BDNF on PC12 cells proliferation. The PC12 cells were seeded at the density of 1×104 cells/cm2 into 96-well plates and 100 µL medium per well. Cells were treated with HAMC-KAFAK/BDNF hydrogels, while the same volume of DMEM was added to the control group. The PC12 cells proliferation was evaluated with a CCK-8 cell proliferation kit (Dojindo Molecular Technologies, Inc., Japan). In brief, at 1, 3, and 7 days, 10 µL CCK-8 solution was applied to each well and incubated for 1 hour, and the absorbance at 450 nm was measured. The experiments were performed in triplicate.
6
Signaling Pathway Modulation in Cell Proliferation
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STATTIC (sc-202818, Santa Cruz Biotechnology, Santa Cruz, CA, USA), SP600125 (s5567) and Anisomycin (A9789) purchased from Sigma (Louis, MO, USA) were dissolved in 100% dimethyl sulfoxide to prepare a 40 mM stock, 20 mM stock and 15 mg/ml stock respectively, and stored at -20°C. Recombinant human interleukin (IL)-6 purchased from R & D Systems (206IL, Minneapolis, MN, USA) was reconstituted in sterile PBS containing 0.1% bovine serum albumen to prepare a 10 μg/ml stock and stored at -20°C. The stock solution was added in culture medium to achieve the indicated final concentrations for cell culture.
The cell proliferation was examined using a CCK-8 cell proliferation kit (Dojindo Laboratories, Kumamoto, Japan), according to the instruction from the supplier. Absorbance was measured at 450 nm with Microplate Reader (Bio-Rad, La Jolla, CA, USA). After 24 hr serum depletion (0.2% FBS), cells were subsequently incubated in 10% FBS medium for an additional 24 hr before harvest. Cell cycle assessment and data analysis were carried out referring to our previous report
[6 (link)] using FACS Calibur flow cytometry (Becton Dickinson, Franklin Lakes, NJ, USA) equipped with the ModiFit LT v2.0 software.
The cell proliferation was examined using a CCK-8 cell proliferation kit (Dojindo Laboratories, Kumamoto, Japan), according to the instruction from the supplier. Absorbance was measured at 450 nm with Microplate Reader (Bio-Rad, La Jolla, CA, USA). After 24 hr serum depletion (0.2% FBS), cells were subsequently incubated in 10% FBS medium for an additional 24 hr before harvest. Cell cycle assessment and data analysis were carried out referring to our previous report
[6 (link)] using FACS Calibur flow cytometry (Becton Dickinson, Franklin Lakes, NJ, USA) equipped with the ModiFit LT v2.0 software.
7
Cytotoxicity of SPION-Coated Cells
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Cells were cultured in a 96-well plate at 5 × 103 cells/well, and incubated at 37 °C with 5% CO2 atmosphere for 24 hrs. The cell culture medium of each well was then replaced with 150 μL fresh complete medium containing SPIONs clusters@PDA at various Fe concentrations. Meanwhile, the cells incubated with cell culture medium only were prepared as untreated control. After incubation for 24 hrs or 48 hrs at 37 °C, cytotoxicity was performed by using a CCK-8 cell proliferation kit following the manufacture’s protocol (Dojindo Molecular Technologies, Tokyo, Japan). A microplate reader (Spectra Max M5) was used to measure the absorbance of the solution in each well at 450 nm. All experiments were performed in quadruplicate.
8
CHI and BTZ Cytotoxicity Assay
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The indicated cells were planted in the 96-well plates (5 × 104 cells / well) and treated with different concentrations of CHI or combined with BTZ for 48 hours. Cell viability was assessed using the CCK-8 cell proliferation kit according to the manufacturer's instructions (Dojindo Laboratories, Kumamoto, Japan).
9
Cytotoxicity of HA-Tb Nanorods on MC3T3-E1 Cells
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MC3T3-E1 cells were seeded in 96-well microplates at a density of 1 × 105 cells mL−1 and cultured for 2 h, 4 h, one day, and seven days in media containing 25, 50, and 100 μg/mL HA-Tb nanorods. After incubation, cytotoxicity was measured by WST-8 assay using a CCK-8 cell proliferation kit (Dojindo Laboratories, Kamimashiki Gun, Kumamoto, Japan). A Varioskan Flash instrument (Bio-Rad 680, Microplate Master, Hercules, CA, USA) was used to measure absorbance at 450 nm. All experiments were carried out in triplicate, and three independent experiments were performed.
10
Cell Proliferation Assay with CCK-8
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A cell counting kit-8 (CCK-8) cell proliferation kit was purchased from Dojindo Laboratories, (Kumamoto, Japan). All the experimental protocols were conducted in accordance with the manufacturers' instructions. Briefly, cells were seeded into a 96-well plate at 1×103 cells/well and cultured at 37°C. CCK-8 solution was added to each well at the indicated times points and then incubated at 37°C at 0, 12, 24, 36 and 48 h, then for a futher 2 h. The absorbance at 450 nm was measured with a microplate reader (Bio-Rad Laboratories, Inc., Hercules, CA, USA). The experiments were repeated in triplicate and three independent experiments were performed.
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