Cells were resuspended with different concentrations of PA and seeded in 96-well plates at a density of 3 × 103 per well. Cellular cytotoxicity was evaluated using the Cell Counting Kit-8 (CCK-8; Dojindo). After 24 h of treatment, the culture medium was changed to DMEM containing 10 μL per well of Cell Counting Kit-8 (CCK-8, Dojindo) solutions and incubated for a further 2 h at 37°C. The optical density (OD) value was measured at a 490 nm wavelength.
Cell counting kit 8 cck 8
Manufactured by Dojindo Laboratories
Sourced in Japan, United States, China, Germany
The Cell Counting Kit-8 (CCK-8) is a colorimetric assay used to measure the number of viable cells in cell proliferation and cytotoxicity assays. It utilizes a water-soluble tetrazolium salt that is reduced by cellular dehydrogenases, resulting in the formation of a colored formazan dye. The amount of formazan dye is directly proportional to the number of living cells in the culture, which can be quantified by measuring the absorbance of the solution.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (232 mentions)
Microplate reader, by Thermo Fisher Scientific (149 mentions)
Microplate reader, by Bio-Rad (122 mentions)
Microplate reader, by Agilent Technologies (80 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (71 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (67 mentions)
Trypsin, by Thermo Fisher Scientific (40 mentions)
Multiskan fc, by Thermo Fisher Scientific (40 mentions)
Microplate reader, by Molecular Devices (32 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (32 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (29 mentions)
β actin, by Cell Signaling Technology (29 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (28 mentions)
96 well plate, by Corning (27 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (26 mentions)
Streptomycin, by Thermo Fisher Scientific (25 mentions)
Matrigel, by BD (23 mentions)
Trypsin edta, by Thermo Fisher Scientific (22 mentions)
Trizol reagent, by Thermo Fisher Scientific (21 mentions)
Penicillin, by Thermo Fisher Scientific (21 mentions)
3 182 protocols using cell counting kit 8 cck 8
1
Cytotoxicity Evaluation of PA Treatment
2
Cytotoxicity Assessment of ESK-MMAE and Q2L-MMAE
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The cytotoxicity of ESK-MMAE and Q2L-MMAE was assessed on K562-A2-WT1126–134 and A431 cell lines. Briefly, 3000 cells were seeded in 96-well plates and incubated with different concentrations of ESK-MMAE and Q2L-MMAE (3 replicates per concentration, 0 μg/mL was the control group) at 37 °C, 5% CO2 for 96 h. After incubation, cells were incubated with 10% Cell Counting Kit-8 (CCK-8) (Dojindo, Kumamoto, Japan) for 2–3 h, and then the OD450 value was measured using a BioRad Model 680 Microplate Reader. According to the formula: The survival rate = the OD450 value of different concentration wells/the average value of OD450 of the control group × 100%, the survival rate of each well cell was calculated. The IC50, which represents the concentration of a drug that is required for 50% inhibition in vitro, was calculated by GraphPad Prim 6.01 software (Graphpad, San Diego, CA, USA based on the survival rate.
3
Cell Proliferation Assay in MCF-7 Cells
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MCF-7 cells were incubated in 10% Cell Counting Kit-8 (CCK-8) (Dojindo, Kumamoto, Japan) diluted in Dulbecco’s Modified Eagle’s Medium at 37°C until a visual color conversion occurred. Following transfection, cell proliferation rates were determined at 0, 24, 48, 72, 96 hours. A microplate reader (set at 450 nm) was used to measure the absorbance of each well. All experiments were performed in triplicate.
4
Synthesis and Characterization of mPEG-Conjugated Iron Oxide Nanoparticles
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Example 2
Methoxy poly(ethylene glycol)-succinimidyl-succinate (mPEG-SS, MW 2000) was purchased from SunBio, Inc. 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride was purchased from Tokyo Chemical Industry Co., Ltd. Poly-L-lysine hydrobromide (PLL, MW 25,000), hydrocaffeic acid, and rhodamine B isothiocyanate (MW 536.08) were obtained from Sigma Aldrich. The Cell Counting Kit-8 (CCK-8) was purchased from Dojindo Molecular Technologies, Inc. Dialysis membranes were obtained from Spectrum Laboratories, Inc. DMEM, RPMI 1640 medium, Dulbecco's phosphate-buffered saline (PBS), FBS, penicillin/streptomycin, and trypsin were obtained from Gibco-BRL. Iron oxide nanoparticles having a diameter of 10 nm or less were obtained from the National Creative Research Initiative Center for Oxide Nanocrystalline Materials and School of Chemical and Biological Engineering at Seoul National University.
5
Klotho Modulates Angiotensin II-Induced Vascular Remodeling
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Dulbecco’s modified Eagle’s medium (DMEM) was purchased from HyClone (HyClone, Logan, CT, USA). Fetal bovine serum (FBS) was purchased from ABGENT (San Diego, CA, USA). We purchased penicillin, streptomycin, and Angiotensin II from Sigma-Aldrich (St. Louis, MO, USA). The Cell Counting Kit-8 (CCK8) was purchased from Dojindo (Kumamoto, Japan). The rat recombinant Klotho protein was from Cloud-Clone Corporation (Houston, TX, USA). The antibodies used in this study were as follows: mouse monoclonal PCNA (PC10), rabbit polyclonal NF-κB p65 (C-20), rabbit polyclonal p-NF-κB p65 (Ser 536), rabbit polyclonal ERK1/2 (K-23), rabbit polyclonal p-ERK1/2 (Thr202/Tyr204), rabbit polyclonal anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (FL-335) (Santa Cruz Biotechnology, CA, USA); and rabbit polyclonal SM22α (Cloud-Clone Corp., Wuhan, China). Anti-CDK2, anti-CDK4, anti-cyclin D, and anti-cyclin E antibodies were purchased from Wanleibio, Shenyang, China, and anti-Akt and anti-phospho-Akt (Ser473) antibodies were from Cell Signaling (MA, USA). The reactivity with rat antigens is included in all of these antibodies. Through ImageJ software (NIH, USA), the signal intensity was quantified.
6
Cytotoxicity of Bisphenol Analogues
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BPA and BPF were obtained from Sigma-Aldrich (St. Louis, MO, USA), and TMBPF was purchased by Toronto Research Chemicals (Toronto, ON, Canada). Culture media, fetal bovine serum (FBS), and antibiotics were from Gibco (Carlsbad, CA, USA). The cell counting kit-8 (CCK-8) was purchased from Dojindo (Rockville, MD, USA). Recombinant mouse RANKL was from PeproTech (JN, USA). All primary antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA).
7
Leptin-induced Oxidative Stress in BEAS-2B Cells
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BEAS-2B cells (no. ATCC® CRL-9609™) were obtained from the American Type Culture Collection and were tested negative for mycoplasma. DMEM-high glucose, FBS and Trypsin-0.25% EDTA were from Gibco (Thermo Fisher Scientific, Inc.). Recombinant human leptin was purchased from Novoprotein Scientific Inc. Lipopolysaccharide (LPS) was purchased from Sigma-Aldrich (Merck KGaA). mitoQ was purchased from Focus Biomolecules. DMSO and penicillin-streptomycin (pen-strep) were obtained from Solarbio. The Cell Counting Kit-8 (CCK-8) was purchased from Dojindo Molecular Technologies. MitoSOX™ Red mitochondrial superoxide indicator for live-cell imaging (cat. no. M36008), MitoTracker® Red CM-H2XRos (cat. no. M7513) and TRIzol® reagent were purchased from Invitrogen (Thermo Fisher Scientific, Inc.). DAPI staining solution was obtained from Beyotime Institute of Biotechnology, Inc. PrimeScript™ RT Master Mix (cat. no. RR036A) was obtained from Takara Bio, Inc. LightCycler® 480 SYBR-Green I Master Mix (cat. no. 04707516001) was purchased from Roche Diagnostics. ELISA kit (cat. no. ml058059; Shanghai Enzyme-linked Biotechnology Co., Ltd.) and all other chemicals of the highest purity available were purchased from local companies.
8
Icaritin Enhances Osteogenesis in Cells
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Icaritin (>98% purity) was purchased from Shanghai U-sea Biotech Co. Ltd. modified Eagle's medium of alpha (α-MEM) and fetal bovine serum (FBS) were both obtained from Hyclone (Hyclone; GE Healthcare Life Sciences); The Cell Counting Kit-8 (CCK-8) was purchased from Dojindo Molecular Technologies, Inc. Dimethyl sulfoxide (DMSO), trypsin and TRIzol® reagent were purchased from Gibco; Thermo Fisher Scientific, Inc.; the alkaline phosphatase (ALP) activity kit, enhanced chemiluminescent detection reagent and micro-BCA assay kit were obtained from Beyotime Institute of Biotechnology. DKK-1 and ICI182780 were purchased from R&D Systems, Inc. The RNA extraction kit was purchased from Takara Bio, Inc. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) primers were obtained from Invitrogen; Thermo Fisher Scientific, Inc. The antibodies used were obtained from Cell Signaling Technology, Inc., unless otherwise indicated. Other reagents used in the experiment were purchased from Sigma-Aldrich; Merck KGaA. Icaritin was dissolved in DMSO and the final concentration of DMSO was 0.05% (v/v). ICI 182780 was dissolved in DMSO and stored at 4°C. Cells were pretreated with ICI 182780 (1 µM) for 30 min at 37°C, followed by icaritin treatment.
9
Synthesis and Characterization of mPEG-Conjugated Iron Oxide Nanoparticles
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Example 2
Methoxy poly(ethylene glycol)-succinimidyl-succinate (mPEG-SS, MW 2000) was purchased from SunBio, Inc. 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride was purchased from Tokyo Chemical Industry Co., Ltd. Poly-L-lysine hydrobromide (PLL, MW 25,000), hydrocaffeic acid, and rhodamine B isothiocyanate (MW 536.08) were obtained from Sigma Aldrich. The Cell Counting Kit-8 (CCK-8) was purchased from Dojindo Molecular Technologies, Inc. Dialysis membranes were obtained from Spectrum Laboratories, Inc. DMEM, RPMI 1640 medium, Dulbecco's phosphate-buffered saline (PBS), FBS, penicillin/streptomycin, and trypsin were obtained from Gibco-BRL. Iron oxide nanoparticles having a diameter of 10 nm or less were obtained from the National Creative Research Initiative Center for Oxide Nanocrystalline Materials and School of Chemical and Biological Engineering at Seoul National University.
10
Docetaxel-Loaded PLGA Nanoparticles
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CNs (50 mg/mL) were purchased from Lummy (China).
PFH was purchased
from Alfa Aesar (U.K.). PLGA (50:50, 12 000 Da MW) was purchased
from Daigang (China). Docetaxel (DOC, 20 mg) was purchased from Hengrui
Medicine (China). Poly(vinyl alcohol) (PVA 25 000 Da MW) was
obtained from Sigma-Aldrich. Methylene chloride (CH2Cl2) was purchased from Chuandong (China). Isopropyl alcohol
(C3H8O) was purchased from Yangzi (China). The
Cell Counting Kit-8 (CCK-8) was purchased from Dojindo (Japan). Agarose
was purchased from Invitrogen (USA). Annexin V-FITC was purchased
from BioVision. Deionized water was obtained with a Milli-Q Plus system
(Millipore Corporation). Natural saline (NS, 0.9%) was purchased from
Tiansheng (China).
PFH was purchased
from Alfa Aesar (U.K.). PLGA (50:50, 12 000 Da MW) was purchased
from Daigang (China). Docetaxel (DOC, 20 mg) was purchased from Hengrui
Medicine (China). Poly(vinyl alcohol) (PVA 25 000 Da MW) was
obtained from Sigma-Aldrich. Methylene chloride (CH2Cl2) was purchased from Chuandong (China). Isopropyl alcohol
(C3H8O) was purchased from Yangzi (China). The
Cell Counting Kit-8 (CCK-8) was purchased from Dojindo (Japan). Agarose
was purchased from Invitrogen (USA). Annexin V-FITC was purchased
from BioVision. Deionized water was obtained with a Milli-Q Plus system
(Millipore Corporation). Natural saline (NS, 0.9%) was purchased from
Tiansheng (China).
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