CellTox Green cytotoxicity assay (Promega) was employed to evaluate changes in Hla or LukAB cytolysis activity in the presence of HS oligos following manufacturer’s protocol. Due to Hla primarily targeting epithelial and endothelial cells and LukAB primarily targeting leukocytes, A549 and HL-60 cells were used as host target cells respectively. Briefly, A549 cells were seeded at 4×104 cells/well and differentiated PMN-HL60 cells were seeded at 1×105 cells/well in black, clear-bottom 96-well tissue culture plates. A549 cells were treated with Hla (Sigma-Aldrich, 0 to 50nM) and HS oligos (0 or 10 μM) for 24 hours. Changes in A549 cytotoxicities were plated in triplicate and quantified by LDH release (CytoTox 96 Assay, Promega) by measuring absorbance at 490 nm with a microplate reader. Differentiated PMN-HL60 were treated with LukAB (0 to 100nM) and HS oligos (0 or 10 μM) for up to 2 hours at 37°C and 5% CO2. Changes in PMN-HL60 cytotoxicity were plated in triplicate and quantified by membrane integrity (CellTox Green cytotoxicity assay, Promega) by measuring fluorescence was measured at 485/535 nm using a microplate reader.
Celltox green cytotoxicity assay
Manufactured by Promega
Sourced in United States, Switzerland, United Kingdom, Italy
The CellTox Green Cytotoxicity Assay is a fluorescent-based cell viability assay used to determine the cytotoxicity of compounds or other treatments on cells in culture. The assay measures the release of DNA from dead or dying cells, providing a quantitative assessment of cytotoxicity.
Automatically generated - may contain errors
Lab products found in correlation
Realtime glo mt cell viability assay, by Promega (6 mentions)
Celltiter glo 2.0 assay, by Promega (6 mentions)
Celltiter glo luminescent cell viability assay, by Promega (5 mentions)
Incucyte zoom, by Sartorius (4 mentions)
Celltox green dye, by Promega (3 mentions)
Ionomycin, by Merck Group (3 mentions)
Celltiter glo assay, by Promega (3 mentions)
Cytation 3 imaging reader, by Agilent Technologies (3 mentions)
Cellevent caspase 3 7 green detection reagent, by Thermo Fisher Scientific (3 mentions)
Propidium iodide, by Thermo Fisher Scientific (3 mentions)
Glomax, by Promega (2 mentions)
Incucyte zoom real time imager, by Sartorius (2 mentions)
Victor3 1420 multilabel counter, by PerkinElmer (2 mentions)
Cytotox fluor assay, by Promega (2 mentions)
Fluostar optima, by BMG Labtech (2 mentions)
Envision multiplate reader, by PerkinElmer (2 mentions)
Celltiter 96 aqueous mts reagent, by Promega (2 mentions)
Cyquant ldh cytotoxicity assay, by Thermo Fisher Scientific (2 mentions)
Steadyglo reagent, by Promega (2 mentions)
Bca assay, by Thermo Fisher Scientific (2 mentions)
119 protocols using celltox green cytotoxicity assay
1
Cytotoxicity Assays for Hla and LukAB
2
Evaluating Artificial Mucin-Mimicking Glycopolymer Cytotoxicity
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HCEs were seeded at a concentration of 5 × 103 cells/well in 96-well plates and allowed 4 h for surface attachment. Afterwards, a medium with different artificial mucin-mimicking glycopolymers (in the 1%, 0.1%, and 0.01% final w/v concentration) was added together with the reagents for cytotoxicity/viability assays according to the manufacturer’s instructions. Measurement of changes in membrane integrity as a result of cell death was performed by CellTox™ Green Cytotoxicity Assay (Promega, Madison, WI, USA) and the determination of the number of viable cells was measured via RealTime-Glo™ MT Cell Viability Assay (Promega). Both assays were used together in each well and real-time repeatedly measured at certain time points (0, 1, 2, 18, 24, 42, and 48 h). Cytotoxicity was determined via increased fluorescence and viability according to measurements of luminescence using a GloMax® Discover Microplate Reader (Promega). Cells cultured without glycopolymers were used as control and the HCEs co-cultured with PEI (MW 750k) as negative control (NC). The blank was subtracted for each concentration and type of glycopolymers individually.
Cell growth, confluence, and cell morphology were evaluated during the culture under an inverted microscope (VWR Visiscope IT405H; VWR, Radnor, PA, USA) and images were taken after 1, 24, and 48 h (images after 1 and 24 h are not shown).
Cell growth, confluence, and cell morphology were evaluated during the culture under an inverted microscope (VWR Visiscope IT405H; VWR, Radnor, PA, USA) and images were taken after 1, 24, and 48 h (images after 1 and 24 h are not shown).
3
Cytotoxicity Assessment of Essential Oils
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For the detection of cytotoxicity, the CellTox™ Green Cytotoxicity Assay (Catalog number G8741, Promega, Madison, WI, USA) was used. RAW264.7 were cultured in triplicate at 5 × 103 cells per well into white-walled, opaque assay 96 well plates; 24h post-seeding, cells were treated for 24 and 48 h with different concentrations of LEO or Cfr-LEO (0.005%, 0.01%, 0.02%, 0.05%). THP-1 M0 cells were seeded in triplicate at 1 × 104 cells per well into white-walled, opaque assay 96 well plates and treated for 24 and 48 h with LEO or Cfr-LEO (0.005%, 0.01%, 0.02%, 0.05%). Changes in membrane integrity that occur as a result of cell death were measured by the CellTox™ Green Cytotoxicity Assay following the manufactures instructions. The fluorescence, proportional to cytotoxicity, was measured by Glomax (Promega).
4
Investigating HEI-OC1 Hair Cell Line
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cochlear hair cell line HEI-OC1 [23 (link)] was a generous gift of Dr. Kalinec (House Ear Institute, Los Angeles, CA, USA). High-glucose Dulbecco's Modified Essential Medium (DMEM, cat no. 11965–092) and fetal bovine serum (cat no. 26140–111) were purchased from Life Technologies. Cisplatin (cat no. 479306) and Mission shRNA containing lentiviral particles were purchased from Sigma-Aldrich. ATPlite one step assay (cat no. 6016731) was purchased from Perkin Elmer and CellTox Green Cytotoxicity Assay (cat no. G8741) from Promega.
5
Cell Cytotoxicity and Apoptosis Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The CellTox Green Cytotoxicity Assay (Promega, Milan, Italy) was used to test cell death in CmPC cells treated with 1 μM DOXO for increasing period of times (6, 24, 48 h, Figure 3a ). This assay measures changes in membrane integrity that occur as a result of cell death, using a dye that is excluded from viable cells but preferentially stains the DNA from dead cells. When the dye binds DNA released from cells, its fluorescence properties are substantially enhanced. Therefore, the fluorescence signal produced by the interaction with DNA from dead cells is proportional to cytotoxicity.
Apoptosis/necrosis in CmPC migration conditions (Figure 3b ) were assessed by using a cell death detection ELISA kit (Roche Diagnostics, Basel, Switzerland). Quantification of histone-complexed DNA fragments (mono- and oligonucleosomes) was performed by one-step sandwich immunoassay, measuring nucleosome-bound DNA fragments by photometric analysis.
Apoptosis/necrosis in CmPC migration conditions (
6
Evaluating Cell Toxicity with P[5]a
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A549 cells were grown as described above. The samples were prepared in biological triplicates, and the average and standard deviation shown. Before the experiment, the cells were treated with TrypLE (Thermo Fisher Scientific), split, counted, and aliquoted into 96-well plates at 50,000 mammalian cells per well. Cells were grown for 16 h at 37 °C in DMEM with the following additives: 10% fetal bovine serum (Gibco), 1× PenStep (Sigma) and 1× GlutaMAX (Gibco). The cells were then washed with 37 °C PBS, and 50 µL of “master mix” was added to each well for each condition. The master mix contained 450 µl DMEM medium without any additives, CellTox Green Dye (CellTox™ Green Cytotoxicity Assay, Promega), either 2.5 mM or 100 µM of P[5]a. The fluorescence was read with a Hidex Sense (Hidex) plate reader with 490 nm excitation source and a 520 nm emission filter. Cells were incubated for 16 h at 37 °C. The final results were based on the difference in fluorescence between 2 h and 5 h measurement time points. The linear range and sensitivity of the experiment was determined using DMSO as the toxic agent.
7
Membrane Permeabilization of E. coli by PhciCath5
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Membrane permeabilisation of E. coli ATCC25922 by PhciCath5 was assessed using the Promega CellTox green cytotoxicity assay. E.coli ATCC25922 was sub-cultured onto TSA II blood agar and incubated at 35oc for 24 hours prior to the test. A bacterial suspension was prepared in RPMI to give an OD600 reading of 0.2. PhciCath5 was dissolved in water for cell culture containing 0.01% acetic acid and serial two-fold dilutions prepared in a black 384-well polypropylene plate. The plate was then innoculated with E. coli ATCC25922, producing a total well volume of 30uL and final peptide concentration of 50 to 0.05uM. Fluorescence was then measured at 512nm using the Perkin Elmer Envision multilabel plate reader (0 hours). The plate was then incubated at room temperature and additional fluorescence measurements were recorded at 1, 2, 3 and 4hrs. Membrane permeability was calculated as a percentage relative to the “no inhibitor” control. PhciCath5 concentration which resulted in greater than or equal to 5% E. coli ATCC25922 membrane permeability was reported.
8
CellTox Green Cytotoxicity Assay
9
Evaluating sEV Cytotoxicity on Hepatocytes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To evaluate whether sEVs can have a toxic effect on hepatocyte growth, the CellTox™ Green Cytotoxicity Assay was performed (G8741, Promega, Madison, WI, USA). THLE-2 cells were cultured in triplicate at 5 × 103 cells/well in white-walled, opaque 96-well plates; 24 h post-seeding, cells were treated for 24 and 48 h with approximately 1.5*E10 particles of SW480_sEVs, SW620_sEVs, CRC_P/sEVs (P1-5) and HS/sEVs (HS1-5). Changes in membrane integrity that occur as a result of cell death were measured as relative fluorescence units (RFUs) by Glomax (Promega, Madison, WI, USA).
10
Cell Viability and Cytotoxicity Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell viability and cytotoxicity were determined using RealTime-Glo MT Cell Viability Assay (Promega) and CellTox Green Cytotoxicity Assay (Promega) according to the manufacturer’s protocol. Per well of white opaque 96-well plates (Corning) 1 × 103 cells were seeded. Assay performance was validated with 10 µM ionomycin (Sigma) (Supplementary Fig. 1a, b).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!