AML cells were seeded into a 96-well plate at a density of 20,000 cells/well for cell viability assay. Cells were treated with different Centrinone concentrations or the vehicle DMSO for 24–96 h, then 10 μL/well Cell Counting Kit-8 (CCK-8; Absin, China) was added to the medium and incubated at 37°C for 3 h, followed by measuring absorbance at 450 nm by Spectra Max M2 Microplate Reader (Molecular Devices, United States).
Cell counting kit 8 cck 8
Manufactured by Absin
Sourced in China
The Cell Counting Kit-8 (CCK-8) is a colorimetric assay for the determination of cell viability and cytotoxicity. 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 is directly proportional to the number of living cells.
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Lab products found in correlation
Spectramax m2 microplate reader, by Molecular Devices (1 mentions)
Pluronic f 127 f 127, by Merck Group (1 mentions)
Melatonin, by Merck Group (1 mentions)
Mitochondria isolation kit, by Nanjing Jiancheng (1 mentions)
3 typ, by MedChemExpress (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
Streptozotocin (stz), by Merck Group (1 mentions)
Ros assay kit, by Beyotime (1 mentions)
Annexin 5 fitc apoptosis detection kit, by Keygen Biotech (1 mentions)
Ly294002, by MedChemExpress (1 mentions)
Mitochondrial membrane potential assay kit with jc 1, by Beyotime (1 mentions)
Multiskan fc microplate photometer, by Thermo Fisher Scientific (1 mentions)
Rhodamine conjugated phalloidin, by Thermo Fisher Scientific (1 mentions)
Cathepsin k, by Santa Cruz Biotechnology (1 mentions)
Recombinant rankl and m csf, by R&D Systems (1 mentions)
C fos, by Abcam (1 mentions)
Nfatc1, by Santa Cruz Biotechnology (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
α mem, by Thermo Fisher Scientific (1 mentions)
β actin, by Santa Cruz Biotechnology (1 mentions)
9 protocols using cell counting kit 8 cck 8
1
Cell Viability Assay for AML Cells
2
Transdermal Delivery of Anti-Inflammatory Agents
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1,3,5-Trimethylbenzene (TMB, Mw 120.19 g/mol, AR, 97%), Tris (hydroxymethyl) aminomethane (TRIS, Mw 121.14 g/mol, ≥99.9%), dopamine hydrochloride (Mw 189.64 g/mol, 98%), Triamcinolone acetonide (TA), Lipopolysaccharides from Escherichia coli O55:B5 (LPS) and phenol were purchased from Aladdin (Shanghai, China). Pluronic® F-127 (F127) was purchased from Sigma (Shanghai, China). Bletilla striata polysaccharide (BSP, Mw 390–460 kDa, ≥98%) was acquired from Xi’an Zelang Biotechnology (Xi’an, China). Sodium hyaluronic acid (HA, Mw 5–10 kDa) was obtained from Freda Biology Co., Ltd. (Shandong, China). The polydimethylsiloxane mold (PDMS, Shenzhen Thunder Cloud Information Technology Co., Ltd., China) had 225 (15 × 15) MNs, and the height and width of each needle were 700 and 300 μm, respectively. The distance among needles was 600 μm. TRITC-Phalloidin, 4′,6-diamidino-2-phenyllindole (DAPI) was purchased from Solarbio. Cell Counting Kit-8 (CCK-8) was purchased from Absin Bioscience Inc (Shanghai, China). Human Tumor Necrosis Factor Alpha (TNF-α) ELISA Kit was purchased from Elabscience Biotechnology Co., Ltd. (Wuhan, China). Methanol was the chromatographic grade for chromatographic analysis when High-performance liquid phase (HPLC) was used, and other chemical reagents were analytical grade, both provided by Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China), and used directly.
3
Cell Proliferation Assay with CCK-8
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The Cell Counting Kit-8 (CCK-8) (Absin, Shanghai, China) was used to determine the CC cell proliferation rate. Transfected cells were inoculated into a 96-well plate at a density of 3000 cells/well. After 24 h in culture, 100 μL of serum-free medium containing 10% CCK-8 reagent was added to each well. A plate reader (AutoBio, Zhengzhou, China) was used to measure the absorbance at 450 nm, 2 h after incubation.
4
Melatonin Mitigates Oxidative Stress-Induced Apoptosis
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Melatonin, streptozotocin (STZ), and 2,3,5-triphenyl tetrazolium chloride (TTC) were obtained from Sigma-Aldrich (St. Louis, MO, USA). LY294002 and 3-TYP were purchased from MedChemExpress (MCE; Monmouth Junction, NJ, USA). Dulbecco’s Modified Eagle Medium (DMEM) and penicillin/streptomycin were obtained from GENOM (Hangzhou, China), fetal bovine serum (FBS) were bought from TIANHANG (Zhejiang, China). The mitochondrial membrane potential assay kit with JC-1 and the ROS Assay Kit were got from Beyotime (Shanghai, China). Cell Counting Kit-8 (CCK-8) was obtained from Absin (Shanghai, China). Kits for detecting malondialdehyde (MDA) content, superoxide dismutase (SOD) activity, catalase (CAT) activity, and ATP content and Mitochondria Isolation Kit were purchased from the Nanjing Jiancheng (Nanjing, Jiangsu, China). The Annexin V-FITC Apoptosis Detection Kit was obtained from KeyGEN (Nanjing, Jiangsu, China). Primary antibodies against p-Akt, Akt, cytochrome c (cyto-C), and β-actin, as well as the secondary antibodies, were all purchased from Cell Signaling Technology (CST; Boston, MA, USA). Primary antibodies against SIRT3, SOD2 (acetyl K68) (ac-SOD2), SOD2, TFAM, TFAM were purchased from Abcam (Cambridge, MA, USA).
5
Cell Proliferation Assay Protocol
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The GBC-SD and NOZ cells were cultured in a 96-well plate at a density of 5,000 cells/well for 0, 24, 48, and 72 hours. The cultured medium in each well was exchanged with fresh medium containing 10% Cell Counting Kit-8 (CCK-8; Absin Bioscience, Inc., Shanghai, China) and incubated at 37 ℃ for 2 hours. The optical density (OD) values of the medium were measured at 450 nm using a MultiskanTM FC Microplate Photometer (Thermo Fisher Scientific, USA).
6
Osteoclastogenesis Assay Protocol
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GA (PubChem CID: 370) was supplied from FeiyuBio (Nantong, China) and dissolved into a storage concentration of 100mM with dimethyl sulfoxide (DMSO). FBS and α-MEM were purchased from Gibco (Rockville, MD, United States) and AusGeneX (Brisbane, Australia), respectively. Recombinant RANKL and M-CSF were provided by R&D (R&D Systems, MN, United States). Cell Counting Kit-8 (CCK-8) was supplied by Absin (Shanghai, China). Rhodamine‐conjugated phalloidin were purchased from Thermo (San Jose, CA, USA). The Cell Signaling Technology (Beverly, MA, USA) supplied the primary antibodies against p-AKT, AKT, p-ERK, p-JNK, JNK, p‐P38, and P38. The antibodies against β-Actin, NFATc1, Cathepsin K (CTSK), and ERK were obtained from Santa Cruz (San Jose, CA, USA). C-Fos were obtained from Abcam (Cambridge, MA, USA). RAW264.7 cells were kindly provided by Stem Cell Bank, Chinese Academy of Sciences. Sterile bone slices were obtained from IDS (London, UK).
7
Cell Viability Assay with CCK-8
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Cell viability was estimated using the Cell Counting Kit-8 (CCK-8; absin, Shanghai, China). The IPEC-J2 cells at the logarithmic stage were placed in 96-well plates for transfection and CPB2 toxin treatment. After 24 h, 10 μL of CCK-8 reagent was added per well and fostered for 2 h. The absorbance was read at 450 nm through a multifunctional enzyme marker (Molecular Devices, Silicon Valley, CA, USA).
8
Chemokine Receptor Antagonist Effects on RA-FLS Viability
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RA-FLS were seeded into 96-well plates at a density of 5x103 cells/well for 24 h and subsequently treated with different concentrations of several chemokine receptor antagonists: BX471 (50, 100 and 150 nM; CCR1 antagonist, Sigma-Aldrich; Merck KGaA), AMG487 (0.5, 1 and 2 µM; CXCR3 antagonist, Sigma-Aldrich; Merck KGaA) and SB225002 (0.1, 0.2 and 0.4 µM; CXCR2 antagonist, Sigma-Aldrich; Merck KGaA). Following incubation for 24 h at 37˚C, 10 µl Cell Counting Kit-8 (CCK-8; Absin Bioscience, Inc.) reagent was added to each well and the optical density was measured at a wavelength of 450 nm.
9
Cell Viability Assay for Hepatocellular Carcinoma
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The compounds were tested for HCC cell lines using the Cell Counting Kit-8 (CCK-8) (Absin, Shanghai, China). The human HCC cell lines Huh7 and HepG2 were cultured in high-glucose Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 1% penicillin, and 1% penicillin/streptomycin at 37 °C with 5% CO2. Huh7 and HepG2 cells were seeded into white 96-well plates at 3 × 103 cells per well in 100 μL suspension and pre-incubated overnight. The cells were then treated with different compound concentrations and incubated for 48 h. Control wells just contained cell suspension, while blank wells were added with complete media. The effects on proliferation were determined by the addition of 10 µL CCK-8 reagent, followed by incubation for 1 h. Finally, the absorbance of the reaction was measured at 450 nm using a Victor Nivo Multimode Microplate Reader. Cell viability was calculated as follows:
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