The assessment of cell viability in H9c2 cells was carried out with the adoption of CCK-8 method [19 (link)]. H9c2 cells treated accordingly in each group were inoculated in 96-well plates at 37°C in 5% CO2 for 24 h. Next, each well was added with 10 μL of CCK-8 solution (GlpBio, Montclair, CA, USA) using a repetitive pipette and incubated for 2 h, strictly comply with the manufacturer’s notes. Determination of absorbance in each well at 450 nm was performed utilizing an enzyme marker (Molecular Devices, Shanghai, China).
Enzyme marker
Manufactured by Molecular Devices
Sourced in United States, China, Germany
The Enzyme Marker is a laboratory instrument designed to detect and quantify the presence of specific enzymes in a sample. It utilizes advanced detection methods to provide accurate and reliable measurements of enzyme activity. The core function of the Enzyme Marker is to enable researchers and scientists to analyze and study the behavior of enzymes in various applications.
Automatically generated - may contain errors
Lab products found in correlation
Cck 8 solution, by GLPBIO (1 mentions)
Cck 8 reagent, by Biosharp (1 mentions)
Cck 8, by Thermo Fisher Scientific (1 mentions)
Cell counting kit 8 (cck8), by Thermo Fisher Scientific (1 mentions)
Eclipse ts2r, by Nikon (1 mentions)
Cck 8, by Dojindo Laboratories (1 mentions)
Highpure maxi plasmid kit, by Tiangen Biotech (1 mentions)
10 protocols using enzyme marker
1
Assessing Cell Viability in H9c2 Cells
2
CCK-8 Assay for Cell Viability
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The CCK-8 assay was used to detect cell viability. HK-2 cells at logarithmic growth stage were taken and inoculated in 96-well plates at a cell density of 4 × 104/mL. Then, the plates were placed in a constant temperature incubator set at 37 °C and 5% CO2. The supernatant was discarded, and new medium (100 µL) containing CCK8 reagent (10 µL) (Biosharp, China) was added; the medium was gently shaken several times and incubated at 37 °C and 5% CO2 for 2 h. The absorbance value of each well was measured at 450 nm using an enzyme marker (molecular devices, Germany).
3
Evaluating E-D Compounds on SW620 Cell Viability
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The effects of E-D1~E-D4 and compounds 1 –7 on the cell viability of SW620 were assayed using MTT powder. Cells were inoculated at 8 × 103 cells/well in 96-well plates overnight and treated with drugs for 24 h. Then, cells were incubated with MTT reagent at 37 °C for 4 h. The supernatant was gently discarded and DMSO was added for lysis. Cell viability was calculated by measuring the absorbance at 490 nm using an enzyme marker (Molecular Devices, Shanghai, China).
4
Cell Viability Assay Using CCK-8
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In this study, the viability of BCAP-37 cells was assayed using the Cell Counting Kit-8 (CCK8) (IC-1519, InCellGene, Tx. USA) method. Cells were inoculated in 96-well cell culture plates according to 1500 cells/well, followed by siRNA transfection. After transfection was completed, the cells were continued to be incubated in the incubator, and 10ul of CCK-8 reagent was added at the same time every day and assayed at 0 h, 24 h, 48 h, and 72 h, respectively. Finally, the absorbance at 450 nm was detected using an enzyme marker (Molecular Devices, USA).
5
Cell Proliferation Assay Protocol
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Each experimental group digested and resuspended in full culture media. Cell proliferation was measured at 1d, 2d, 3d, and 4d using the CCK-8 (Invitrogen, USA). In 450 nm, optical densities were measured using an enzyme marker (Molecular Devices, Rockford, IL).
6
Papillary Thyroid Cancer Cell Proliferation
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Cells from each experimental group in the logarithmic growth phase and in good growth conditions were digested and resuspended in full culture media. Cells were seeded at 3000 papillary thyroid cancer cells per well in 96-well plates was measured using the Cell Counting Kit-8 (Invitrogen, USA) according to the manufacturer’s instructions at 24 h, 48 h, 72 h, and 96 h. The optical density values at 450 nm were measured using an enzyme marker (Molecular Devices, Rockford, IL, USA).
7
Biocompatibility Evaluation of Microspheres and Nanoparticles
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Caco‐2 cells were selected to study the biocompatibility of microspheres. Briefly, Caco‐2 cells were seeded in 24‐well plates (1×104 cells per well) and cocultured with the various microsphere extracts at 37 °C under 5% CO2, similar to the previous reports.[34 ] On days 1, 3, and 5, the cells were incubated with Calcein AM/PI buffer (Proteintech Co., Ltd., China) for 15 min and subsequently observed under a fluorescent microscope (Nikon ECLIPSE Ts2R, Japan). Caco‐2 cells were seeded in 96‐well plates (1 × 104 cells per well) and cocultured with the various microsphere extracts at 37 °C under 5% CO2. After cultivation for 1, 3, and 5 days, it was incubated for 1 h in the medium containing a 10% CCK‐8 (Dojindo, Japan) before the absorbance measurement at 450 nm by an enzyme marker (Molecular Devices, Japan).
BMMs and HUVECs were used to investigate the biocompatibility of PDAP NPs. BMMs or HUVECs were seeded in 96‐well plates (1×104 cells per well) and coculture with the different concentrations of PDAP NPs (0, 12.5, 25, and 50 µm ) at 37 °C under 5% CO2. After cultivation for 48 h, the medium containing 10% CCK‐8 was added and further incubated it for 1 h, and detected its absorbance at 450 nm by enzyme marker.
BMMs and HUVECs were used to investigate the biocompatibility of PDAP NPs. BMMs or HUVECs were seeded in 96‐well plates (1×104 cells per well) and coculture with the different concentrations of PDAP NPs (0, 12.5, 25, and 50 µ
8
Cytotoxicity Evaluation of CaF2:Y,Gd,Nd NPs
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In vitro cytotoxicity of CaF2:Y,Gd,Nd NPs was studied on 4T1 cells. The 4T1 cells (1 × 104) were inoculated in 96-well plates, 100 μL of medium was added to each well, and the cells were allowed to adhere overnight. The medium was then replaced with DMEM containing different concentrations of CaF2:Y,Gd,Nd NPs (2000, 1000, 500, 250, 125, 62.5, and 31.25 µg/mL). The medium was removed at the indicated intervals (24, 48, and 72 h) and cell viability was assessed by MTS (Promega, Madison, WI, USA) according to the manufacturer’s instructions. The absorbance (OD) was measured at 490 nm using an enzyme marker (Molecular Devices, San Jose, CA, USA). Data are expressed as a percentage of the experimental group versus the control group. The experiment was performed in quadruplicate.
9
Evaluating Cell Viability with CCK-8
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The effect of SS on the viability of SW620, SW480, A549 and MGC803 cells was assayed using CCK-8 (Invigentech, Shanghai, China). Cells (8 × 103) were inoculated in 96-well plates overnight and treated with drugs for 24 h. Then, cells were incubated with CCK-8 for 1 h at 37 °C. Cell viability was calculated by measuring absorbance at 450 nm with an enzyme marker (Molecular Devices, Ismaning, Germany).
10
Transcription Factor Binding Site Analysis
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SY5Y human neuroblastoma cells (kindly provided by Dr. Chunhua Chen, Peking University) were used. SY5Y cells were cultured in DMEM supplemented with 10% FBS and antibiotics. Candidate sequences were obtained by CUT&Tag. Two pCDNA3.1 (+) plasmids expressing the transcription factors ESR1 and ESR2 and six PGL3-promoter plasmids expressing the binding sites of interest were constructed. The original luciferase plasmid with only TK was used as the control. All the constructed sequences and the sequences of the primers used are listed in Supplementary Table 4 . The plasmids were extracted with a HighPure Maxi Plasmid Kit (Tiangen, DP116). The cells were then cotransfected with pRL-TK (as a reference plasmid), pCDNA3.1 (+) and the pGL3 promoter. Luciferase assays were performed with three independent groups of transfected cells using a Dual-Glo Luciferase Assay Kit from Genomeditech. The absorbance was measured by an enzyme marker (Molecular Devices), and the Luc/Rena value was calculated for each plasmid. The original luciferase plasmid with only TK was used as the control. The TLuc/Rena values of the plasmid-expressed transcription factor and promoter action site sequences were measured and calculated as luciferase activity.
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