Cells were seeded in a 96-well plate at a density of 1×103 cells/well, and the cell viability was then assessed at different time points (0 hr, 24 hrs, 48 hrs, 72 hrs, 96 hrs and 120 hrs) by CCK-8 kit (Transgen, China). Each well was added with 10 μl of CCK-8 and 90 μl of fresh serum-free medium, followed by incubation for 4 hrs at room temperature. Afterward, we performed a Microplate Reader at 450 nm to measure the absorbance levels of samples.
Cck 8 kit
Manufactured by Transgene
Sourced in China
The CCK-8 kit is a colorimetric assay for the quantitative determination of cell viability and cytotoxicity. The kit utilizes the tetrazolium salt WST-8 to produce a water-soluble formazan dye upon bioreduction in the presence of an electron carrier. The amount of formazan dye generated is directly proportional to the number of living cells in the sample, which can be measured spectrophotometrically.
Automatically generated - may contain errors
Lab products found in correlation
Goat anti mouse secondary antibody, by Abcam (1 mentions)
Mg132, by Selleck Chemicals (1 mentions)
Microreader, by Bio-Rad (1 mentions)
1 1 diphenyl 2 picrylhydrazyl dpph, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
2 2 azino bis 3 ethylbenzothiazoline 6 sulfonic acid, by Merck Group (1 mentions)
Dmem medium, by Thermo Fisher Scientific (1 mentions)
9 protocols using cck 8 kit
1
Cell Viability Assay Using CCK-8
2
Nrf2 Regulation in MODE-K Cells
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PC, DMSO (Beijing, Solarbio), RPMI 1640, FBS (Israel BI), CCK-8 kit (Beijing, TransGen Biotech), cycloheximide (CHX) (American Sigma), MG132 (Selleck), and MODE-K cells were purchased from Shanghai Guandao Biotechnology Company. Primary and secondary antibodies used in experiments, specifically anti-Nrf2, anti-Ub, anti-β-actin, anti-HO-1, anti-Keap1, anti-BACH1, anti-NQO1 (US CST), anti-GAPDH (Suzhou Ruiying), goat anti-rabbit, and goat anti-mouse secondary antibodies, were all from Abcam, USA.
3
Cell Viability Assessment Using CCK-8
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Cell viability was detected using the CCK-8 kit, according to the manufacturer’s instructions (TransGen Biotech, China). Briefly, the culture medium was removed and CCK-8 (10%, 100 μl/well) was added to each well and incubated for 1 h. A microplate reader was used to measure the absorbance OD value at 450 nm.
4
Cell Proliferation Assay using CCK-8
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For the CCK-8 assay, ICP-1 cells were inoculated in 96-well plates and transfected with plasmids or siRNA. Cell proliferation and viability were then monitored at 12, 24, 36, and 48 h using the CCK-8 kit (TransGen Biotech, Beijing, China) following the manufacturer’s instructions. This analysis was performed using a microreader (Bio-Rad, Hercules, CA, United States) to measure the absorption value at 450 nm.
5
Schwann Cell Glucose Intervention Assay
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RSC96 rat Schwann cell line was obtained from Procell (Wuhan, China). Cell culture conditions were DMEM (high glucose) + 10% fetal bovine serum (FBS) + 1% penicillin and streptomycin at 37 °C with 5% CO2.
The cell proliferation for glucose intervention was detected using CCK-8 kit (TransGen, Beijing, China) following the manufacturer’s instructions. The OD value was gathered at 450 nm. High glucose model was established by intervening RSC96 cells with 150 mM glucose for 24 h, as shown in Table S1. Cells were randomly divided into six groups: RSC96 (control), high glucose, high glucose + low dose (0.01 mg/ml of CQYG pretreated for 24 h), high glucose + medium dose (0.05 mg/ml of CQYG pretreated for 24 h), high glucose + high dose (0.1 mg/ml of CQYG pretreated for 24 h), high glucose + TMAO (5 mM of CQYG pretreated for 24 h).
The cell proliferation for glucose intervention was detected using CCK-8 kit (TransGen, Beijing, China) following the manufacturer’s instructions. The OD value was gathered at 450 nm. High glucose model was established by intervening RSC96 cells with 150 mM glucose for 24 h, as shown in Table S1. Cells were randomly divided into six groups: RSC96 (control), high glucose, high glucose + low dose (0.01 mg/ml of CQYG pretreated for 24 h), high glucose + medium dose (0.05 mg/ml of CQYG pretreated for 24 h), high glucose + high dose (0.1 mg/ml of CQYG pretreated for 24 h), high glucose + TMAO (5 mM of CQYG pretreated for 24 h).
6
Antioxidant Evaluation of R. griseocarnosa
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The dry R. griseocarnosa sporocarp was purchased from Fujian, Southeast of China. The material was identified by Professor Bau Tolgor of Jilin agriculture university, Changchun, China.
1,1-Diphenyl-2-picrylhydrazyl (DPPH) and 2,2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) were purchased from sigma Co. The Fetal bovine serum (FBS) and DMEM medium were purchased from Gibco, Biomycs was purchased from Biological Industries. CCK8 kit was purchased from TransGen Biotech. Other chemicals including anhydrous ethanol, petroleum ether, phenol, acetone, sulfuric acid, potassium persulfate, H2O2, Trolox, Vitamin C and salicylic acid-ethyl alcol were purchased locally and were of analytical grade.
1,1-Diphenyl-2-picrylhydrazyl (DPPH) and 2,2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) were purchased from sigma Co. The Fetal bovine serum (FBS) and DMEM medium were purchased from Gibco, Biomycs was purchased from Biological Industries. CCK8 kit was purchased from TransGen Biotech. Other chemicals including anhydrous ethanol, petroleum ether, phenol, acetone, sulfuric acid, potassium persulfate, H2O2, Trolox, Vitamin C and salicylic acid-ethyl alcol were purchased locally and were of analytical grade.
7
CCK8 Assay for Cell Viability
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Residual cell viability was measured by CCK8 kit (#FC101, TransGen) according to the manufacturer's instructions.
8
Cell Viability Assay with CCK-8
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Cell viability was determined using CCK-8 Kit (TransGen Biotech, Beijing, China) according to the manufacturer’s protocol. Briefly, cells (100 μL/well, 1 × 104 cells/ml) were seeded on 96-well plates for 24 h, then treated with or without different concentrations of various inhibitors and incubated at 37°C for different times. CCK-8 solution was added to each well of the plates, followed by incubation at 37°C for 1 h. The optical density (OD) was measured at 450 nm using an enzyme-labeled instrument (BMG. FLUO star Optima, Offenburg, Germany). The cell inhibitory rate was calculated by using the following equation:
Cell inhibitory rate = [1 − (OD experiment − OD blank)/(OD control − OD blank)] × 100%.
All experiments were performed in triplicate and repeated three times independently.
Cell inhibitory rate = [1 − (OD experiment − OD blank)/(OD control − OD blank)] × 100%.
All experiments were performed in triplicate and repeated three times independently.
9
Cell Proliferation Assay and HCC Tissue Microarray
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Cell proliferation assay was performed by using a CCK8 kit (Transgene, Beijing, China). Brie y, cells (3×10 3 cells/well) were seeded into 96-well plates and incubated for 1, 2, 3, 4, and 5 days. At the indicated time point, 100 µl fresh medium plus10 µl CCK8 solution was added to each well pre-emptied. The plate was then incubated for additional 2 hours before measuring the absorbance at 450 nm wavelength using a microplate reader (M5e, MD Co. USA). The assays for each time point were performed in triplicate and the experiments were biologically repeated at least three times. Tissue microarrays HCC tissue microarrays (TMA) were purchased from Alenobio (cat# LV2084) (Xi'an, Shaanxi, China). This TMA contained 83 HCCs, 5 cholangiocellular carcinomas, 2 adenosquamous carcinomas, 1 each of mixed carcinoma, carcinoma sarcomatodes, squamous cell carcinoma, and papillary carcinoma, and 10 normal liver tissues. The TMA was generated in duplicate so as to compare the staining in different areas of the same sample. Tumor characteristics of these TMA samples were available including age, gender, tumor type, histological grade, tumor stage, and metastasis status (Supplementary Table S1).
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