A549 and PC9 cells were digested with 0.25% trypsin solution, and the cell density was readjusted to 3 × 104cells/mL. The resuspended cells were seeded into 96-well plates at 200 µL/well. The 96-well plate was placed in the incubator for 24 h, and then the medium was changed to an equal volume of shikonin liquid at different concentrations (1 µM, 2 µM, 3 µM, 4 µM, 5 µM, 6 µM, 7 µM, 8 µM, 9 µM, and 10 µM). After 24 h, the drug solution was discarded, and 20 µL of medium and cell counting kit-8 (CCK-8) solution (Bimake, Shanghai, China) were added to each well to mix and dilute at a ratio of 9:1, and were placed in an incubator for cultivation after 2–4 h. The optical density (OD) value of the CCK-8 solution in the 96-well plate was detected by a microplate reader at 450 nm, and the cell-free well was used as a blank control.
Cck 8 solution
Manufactured by Selleck Chemicals
Sourced in United States, China
The CCK-8 solution is a cell counting kit used for measuring cell proliferation and cytotoxicity. It is a colorimetric assay based on the reduction of tetrazolium salt to a colored formazan product. The amount of the formazan dye generated is directly proportional to the number of living cells in the sample.
Automatically generated - may contain errors
Lab products found in correlation
Hoechst 33342, by RiboBio (4 mentions)
Edu reagent, by RiboBio (4 mentions)
Bx63 automatic intelligent uorescence microscope, by Olympus (3 mentions)
Multiskan mk3, by Thermo Fisher Scientific (1 mentions)
Microplate reader, by Agilent Technologies (1 mentions)
Microplate reader, by Bio-Rad (1 mentions)
Microplate reader, by Thermo Fisher Scientific (1 mentions)
Spectramax id3, by Molecular Devices (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Zhongshan Golden Bridge Biotechnology (1 mentions)
P5250, by Solarbio (1 mentions)
El800, by Agilent Technologies (1 mentions)
Mg 63 cells, by American Type Culture Collection (1 mentions)
Elx800, by Agilent Technologies (1 mentions)
34 protocols using cck 8 solution
1
Evaluating Shikonin Cytotoxicity in Lung Cancer Cells
2
Cell Viability Assay with 5FU and Ivosidenib
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HCT8 or HCT8FU cells were seeded into 96-well plates at a density of 1 × 104 cells/well and treated with the required concentration of 5FU or ivosidenib. After being cultured for 24-96 h, 10 μl of CCK-8 solution (Bimake, Shanghai, China) was added. Absorbance at 450 nm was determined and used to evaluate the viability of the cells.
3
Cell Viability Assay Protocol
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3000 cells/well were plated in 96‐well plates and incubated with indicated time. After incubation, cell counting kit‐8 (CCK‐8) solution (B34304; Bimake, TX, USA) was added and incubated following the manufacturer's instructions. The absorbance of samples was measured at 450 nm using a spectrophotometer (Beckman, USA). Six replicates of each sample were analyzed.
4
Cell Viability Assay using CCK-8
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Cells were seeded into 96-well plate and cultured overnight at 37 °C. After the indicated treatments, 10 µL CCK-8 solution (Bimake, China) was added into each well. After an additional incubation at 37 °C for 2 h, the absorbance was measured at 450 nm using the microplate reader (Dynatech, USA).
5
Cell Viability Assay for HaCaT Cells
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HaCaT cells transfected with pHGF were seeded into 96-well plates at a density of 1.0 × 104 cells/well and incubated overnight. Then 10 μl of CCK-8 solution (B34304, Bimake, USA) was added to the cells 24 h after irradiation, and the plates were incubated at 37°C for 2 h. The absorbance values were measured at 450 nm using the Multiskan MK3 enzyme-labeled instrument (Thermo Fisher Scientific, USA). Cell viability was calculated based on the absorbance values of the sample and the control group.
6
AGE-Induced Cell Viability Assay
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Cell viability in cardiomyocytes and H9c2 cells was detected using cell counting kit 8 (CCK8) following manufacturer’s protocol. Cells were seeded at a density of 1 × 104 cells per well in a 96-well culture plate, and three dosages of AGEs (2 μM, 5 μM, and 10 μM) was added in. After culturing for 24 h, the cells were subsequently incubated with CCK8 solution (Bimake, USA) at 37°C for 2 h. The absorbance was measured at 450 nm with a microplate spectrophotometer.
7
VEGF-B Regulation of MIN6 Cell Proliferation
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Cell counting kit-8 (CCK-8) was used to detect the effects of exogenous VEGF-B and VEGF-B knockdown on the proliferation of MIN6 cells. MIN6 cells were seeded in a 96-well plate at a cell density of 2 × 105/mL. After culture in a cell incubator at 37 °C for 6 h, MIN6 cells were treated with exogenous VEGF-B protein or subjected to knockdown of VEGF-B protein. The original medium was discarded and the cells were washed with PBS buffer. Next, the medium was mixed with CCK-8 solution (Bimake, United States) at a ratio of 9:1 and 100 μL was added to each well. The plates were incubated for another 2 h before they were placed in a microplate reader (Bio Tek, United States) to detect the optical density at 450 nm.
8
Cell Growth Assay for SRP14P124A
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To evaluate the impact of SRP14
P124A on cell growth, cells (3000 cells/well) were seeded in a 96-well plate with 100 μL of complete medium per well. Following a 24-h incubation, 10 μL of CCK-8 solution (Bimake, Houston, USA) was added to each well, and the cells were incubated at 37°C for 1 h. The absorption values were measured at 450 nm with a microplate reader over five consecutive days.
P124A on cell growth, cells (3000 cells/well) were seeded in a 96-well plate with 100 μL of complete medium per well. Following a 24-h incubation, 10 μL of CCK-8 solution (Bimake, Houston, USA) was added to each well, and the cells were incubated at 37°C for 1 h. The absorption values were measured at 450 nm with a microplate reader over five consecutive days.
9
Viability Assay of hNSCs
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1 × 104 cells/well (200 μl cell suspension) HPT-hNSCs were placed into a 96-well plate, and cultured in normal hNSC culture medium or growth factor deprived hNSC culture medium. At 0, 24, and 48 h, 10 μl CCK-8 solution (Bimake, B34302) was added directly to each well and incubated for 3 h. Then the absorbance at 450 nm for each well was measured with a microplate reader.
10
Cell Proliferation Assays: CCK8 and EdU
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Cell proliferation was measured by Cell Counting Kit-8 (CCK8) assays and EdU assays.
For CCK8 assays, cells were seeded in 96-well plates at a density of 2000 cells/well and then were incubated for 12 h to allow cell attachment. Then, 10 µL of CCK-8 solution (Bimake, Cat# B34304) was added to each well on days 1, 2, 3, 4, 5 and 6, which was followed by another 2 h incubation. The optical density was then measured at 450 nm using a microplate reader.
For EdU assays, cells were plated at a density of 20,000/dish in confocal dishes. After 24 h of incubation, cells were treated with EdU reagent (RiboBio, Cat# C10310-1) for 2 h according to the manufacturer’s instructions and then were fixed with 4 % paraformaldehyde. One hundred microliters of 1X Apollo®567 staining reaction solution and Hoechst 33,342 (RiboBio, Cat# C10310-1) was added to each dish and then was incubated for 30 min at room temperature on a decolorization shaker. Cells were then visualized using a BX63 automatic intelligent fluorescence microscope (Olympus, Tokyo, Japan).
For CCK8 assays, cells were seeded in 96-well plates at a density of 2000 cells/well and then were incubated for 12 h to allow cell attachment. Then, 10 µL of CCK-8 solution (Bimake, Cat# B34304) was added to each well on days 1, 2, 3, 4, 5 and 6, which was followed by another 2 h incubation. The optical density was then measured at 450 nm using a microplate reader.
For EdU assays, cells were plated at a density of 20,000/dish in confocal dishes. After 24 h of incubation, cells were treated with EdU reagent (RiboBio, Cat# C10310-1) for 2 h according to the manufacturer’s instructions and then were fixed with 4 % paraformaldehyde. One hundred microliters of 1X Apollo®567 staining reaction solution and Hoechst 33,342 (RiboBio, Cat# C10310-1) was added to each dish and then was incubated for 30 min at room temperature on a decolorization shaker. Cells were then visualized using a BX63 automatic intelligent fluorescence microscope (Olympus, Tokyo, Japan).
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