To determine the cytotoxicity of butyric acid, IncuCyte® Cytotoxicity Assay (Essen BioScience Inc., Ann Arbor, MI) was performed. Briefly, EOL-1 cells were seeded into a 96-well plate at a density of 2 x 104 cells per well. After seeding, the medium was replaced with butyric acid and treated with IncuCyte® Cytotox Green Reagent (Essen BioScience Inc.). The viability of the EOL-1 cells cultured at 37˚C in an atmosphere containing 5% CO2 and 95% humidity was measured at 72 h. Apoptotic EOL-1 cells were stained by IncuCyte® Cytotox Green Reagent and the viability of EOL-1 cells was evaluated by measuring green fluorescence through IncuCyte® software (Essen BioScience Inc.).
Incucyte cytotox green reagent
Manufactured by Sartorius
Sourced in United Kingdom
The IncuCyte Cytotox Green Reagent is a fluorescent dye-based reagent that can be used to detect and quantify cell death in real-time cell imaging and analysis systems. The reagent binds to DNA in cells with compromised cell membranes, providing a quantitative measure of cytotoxicity.
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Lab products found in correlation
Incucyte cytotoxicity assay, by Sartorius (4 mentions)
Incucyte software, by Sartorius (4 mentions)
Incucyte, by Sartorius (2 mentions)
Incucyte caspase 3 7 green, by Sartorius (2 mentions)
Incucyte s3 live cell analysis system, by Sartorius (2 mentions)
Evos m7000, by Thermo Fisher Scientific (1 mentions)
Incucyte zoom system, by Sartorius (1 mentions)
Spectramax plus 384 microplate reader, by Molecular Devices (1 mentions)
Prism 9, by GraphPad (1 mentions)
Woundmaker, by Sartorius (1 mentions)
Pen strep, by Thermo Fisher Scientific (1 mentions)
Pen strep, by Sartorius (1 mentions)
Incucyte s3, by Sartorius (1 mentions)
Annexin 5 green dye, by Sartorius (1 mentions)
Fer 1, by Merck Group (1 mentions)
Caspase 3 7 green apoptosis reagent, by Sartorius (1 mentions)
Deferoxamine dfo, by Merck Group (1 mentions)
Nec 1, by Abcam (1 mentions)
Zvad fmk, by Bachem (1 mentions)
19 protocols using incucyte cytotox green reagent
1
Cytotoxicity Evaluation of Butyric Acid
2
Real-Time Live Cell Cytotoxicity Assay
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Cells infected with NucLight Red Lentivirus were seeded in a 48-well plate at a concentration of 20.000 cells/well. The day after, chemicals were added in media containing IncuCyte® Cytotox Green Reagent (Essen Bioscience®) for loss of cell membrane integrity assessment according to the manufacturer’s protocol [44 (link)]. Immediately after, the plate was put on the IncuCyte® (Essen Bioscience® [Ann Arbor, MI, USA]). Three wells were used per replicate and nine pictures per well were taken every 2 h using a 10× lens for each experiment. IncuCyte® software is used to analyze images for both red nucleus count and green cell dying count.
3
NSAID cytotoxicity in cardiomyocytes
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Immortalized human cardiomyocytes were plated (seeding density 2.5 × 104 cells/cm2) in 10% FBS-supplemented medium w/o phenol red into a 96 black well plate. The next day the culture medium was replaced with 1% FBS-supplemented medium w/o phenol red, this culture medium was daily replaced until the seventh day of culture. Then, the cells were exposed to 100 and 200 μM of NSAIDs in 1% FBS-supplemented medium w/o phenol red, and 250 nM of IncuCyte Cytotox Green Reagent (Essen BioScience) was added in the experimental culture medium for counting dead cells. For the apoptosis detection, 5 μM IncuCyte Caspase-3/7 Green (Essen BioScience) was added in the experimental culture medium for counting caspases activation The plates were placed in IncuCyte device (20 × objective), the cytotoxicity and caspases activation were recorded (three images for well, six replicates) every 3 h by both phase contrast and fluorescence scanning for 24 h at 37 °C and 5% CO2. Images were analysed using the Incucyte ZOOM software and the data were reported as green object count per mm2.
4
Apigenin Cytotoxicity Assessment in Nasal Fibroblasts
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In order to determine the cytotoxicity of apigenin, IncuCyte® Cytotoxicity Assay (Essen BioScience Inc., Ann Arbor, MI) was performed. Nasal fibroblasts of 4 x 105cells/ml were seeded into 96-well tissue culture plates. After seeding, the medium was replaced with apigenin and treated with IncuCyte® Cytotox Green Reagent (Essen BioScience Inc.). The viability of the fibroblasts was measured for 72 hours at 37°C in an atmosphere of 5% CO2 and 95% humidity. Apoptotic fibroblasts were stained by IncuCyte® Cytotox Green Reagent. The viability of fibroblasts was calculated by measuring green fluorescence through IncuCyte® software (Essen BioScience Inc.).
5
Real-time Cell Death Quantification
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The IncuCyte® S3 Live-Cell Analysis System and the IncuCyte® Cytotox Green reagent (Essen BioScience, Hertfordshire, UK), which is a cyanine nucleic acid dye that enables real-time evaluation and quantification of cell death, was used. HeLa cells were seeded into 96-well plates and incubated for 24 h to allow for attachment. Cells were exposed to ESE-15-one and ESE-16, respectively, and the IncuCyte® Cytotox Green reagent (final concentration of 250 nM). The plates were placed in the IncuCyte® S3 Live-Cell Analysis System and two pictures per well were taken every 2 h over a 72-h period.
6
Cell Death Quantification Assay
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Preconfluent cell monolayers grown in 24 well plates were infected with the WT or the VP5-KO virus. After virus adsorption, monolayers were gently washed and covered with fresh medium supplemented with 2% FCS and IncuCyte Cytotox Green reagent (Essen Bioscience, Inc., Ann Arbor, MI) following manufacturer’s instructions, and placed within an IncuCyte ZOOM System apparatus. Cultures were monitored every 30 min using a 10x objective lens. Four randomly selected monolayer locations were imaged in each well over the entire time course. IncuCyte Zoom software was used to automatically score and quantify cell death. Presented data corresponds to three independent infections involving the quantification of a total of 12 video recordings.
7
Cytotoxicity Evaluation of Ladarixin
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3T3-L1 were plated (seeding density 1 × 105 cells/cm2) into a 24-wells plate and differentiated as described above. Then, the cells were exposed to ladarixin treatments (10–25 µM) and different concentrations of FBS, and 250 nM of IncuCyte Cytotox Green Reagent (Essen BioScience, Newark, UK) were added in the experimental culture medium for counting dead cells. The plates were put in IncuCyte device (20 × objectives). The cytotoxicity activation was recorded (three images for well) every 3 h by both fluorescence scanning and phase contrast for 72 h at 37 °C and 5% CO2. Images were analyzed utilizing the Incucyte ZOOM software (Newark, UK), and the data were reported as green object count (per image).
8
IncuCyte Cytotox Green Cell Death Assay
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Cell death assay was performed with IncuCyte® Cytotox Green Reagent (4633, Essen Bioscience) according to the manufacturer’s instructions. A final concentration of 1:4000 dilution in culturing medium was used. Cells were imaged by IncuCyte Zoom System (Essen Bioscience) and EVOS M7000 auto digital microscope (Thermo Fisher Scientific) applying Z-stack.
9
Cytotoxicity Evaluation of Peptides
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An IncuCyte cytotoxicity assay (Essen BioScience, Inc., Ann Arbor, MI, USA) was performed to evaluate time-dependent cytotoxicity. RAW 264.7 cells (2 × 105 cells well−1) were seeded into 96-well culture plates and then treated with peptides and IncuCyte Cytotox green reagent (Essen BioScience, Inc.). Up to 200 μM PN5 and 25 μM melittin were used to treat RAW 264.7 cells, with the concentrations serially diluted by half. Cell viability was observed for 48 h at 37°C in a humidified atmosphere of 5% CO2. The cells were stained with IncuCyte Cytotox green reagent. The viability was measured using IncuCyte software (Essen BioScience, Inc.) (56 ).
10
Cytotoxicity Evaluation of Peptides
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An IncuCyte cytotoxicity assay (Essen BioScience, Inc., Ann Arbor, MI, USA) was performed to evaluate time-dependent cytotoxicity. RAW 264.7 cells (2 × 105 cells well−1) were seeded into 96-well culture plates and then treated with peptides and IncuCyte Cytotox green reagent (Essen BioScience, Inc.). Up to 200 μM PN5 and 25 μM melittin were used to treat RAW 264.7 cells, with the concentrations serially diluted by half. Cell viability was observed for 48 h at 37°C in a humidified atmosphere of 5% CO2. The cells were stained with IncuCyte Cytotox green reagent. The viability was measured using IncuCyte software (Essen BioScience, Inc.) (56 ).
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