NRVMs were seeded on 96-well plates (1500 cells/well). After 24 h, the cells were treated with different concentrations of TAG-23, and cell viability was measured by the CCK8 assay using a cell viability assay kit (Promega) according to the manufacturer’s protocols.
Cell viability assay kit
Manufactured by Promega
Sourced in United States
The Cell Viability Assay Kit is a quantitative colorimetric assay for determining the number of viable cells in a sample. The assay is based on the metabolic reduction of a tetrazolium salt, which results in the formation of a colored formazan product. The amount of formazan produced is directly proportional to the number of viable cells, which can be measured spectrophotometrically.
Automatically generated - may contain errors
Lab products found in correlation
Microplate fluorometer, by Thermo Fisher Scientific (1 mentions)
Fluostar optima, by BMG Labtech (1 mentions)
Crystal violet, by Merck Group (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
N acetyl cysteine (nac), by Merck Group (1 mentions)
Reduced glutathione assay kit, by Nanjing Jiancheng (1 mentions)
Automatic microplate reader, by Agilent Technologies (1 mentions)
9 protocols using cell viability assay kit
1
TAG-23 Dose-Dependent Cell Viability
2
Cell Viability Assay for MPP+ and VPA
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MPP+- and VPA-treatment effect on cell viability was assessed using a cell viability assay kit (Promega, Madison, WI, USA), according to the manufacturer's instructions. The cell viability assay uses the indicator dye resazurin to measure the metabolic capacity of cells and estimate the number of viable cells. After 48 h, 25 µL of treated cell samples was collected, and then 5 µL of cell viability assay reagent per well was added and the plates were incubated at 37°C for 1 h. The fluorescent signal was recorded by a microplate fluorometer (Thermo Fisher Scientific Inc.) at 560/590 nm.
3
Characterization of M. ulcerans and S. aureus
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A well-characterized clinical isolate, M. ulcerans 1615 was used in all studies [26] (link). M. ulcerans 1615 were grown in Middlebrook 7H9 liquid media and on Middlebrook 7H10 agar supplemented with 10% oleic acid-albumin-dextrose enrichment (OADC) and incubated at 30°C to reach exponential phase of growth. Bacterial viability was validated by staining using a cell viability assay kit (Promega, Madison, WI, USA). Staphylococcus aureus 502a (ATCC# 27217) was obtained from American Type Culture Collection, cultured on nutrient agar and incubated at 37°C. This strain of S. aureus was isolated from the nares of a nurse, and described in ATCC as being coagulase positive, penicillin sensitive, and sensitive to 10 mcg of tetracycline.
4
Cell Viability Assay Optimization
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Cells were plated in 96-well plates at 1 × 103 cells/well and cultured for 0, 24, 48, 72, 96, and 120 h. The plates were examined using a cell viability assay kit (Promega, Madison, WI, USA) in accordance with the manufacturer's protocols, as described previously 16 (link).
5
Cell Viability Quantification Assay
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Cell viability was quantitated using a Cell viability assay kit (Promega, NSW, Australia). Briefly, Cells were seeded at 5,000 cells per well onto 96-well culture plates and allowed to grow for 24 h followed by desired drug treatments. Post 48 hour treatment, cells were incubated with CellTiter-Blue® reagent for 1 hour at 37°C. The fluorescent signal was recorded using the FLUOstar OPTIMA (BMG Labtech, VIC, Australia).
6
Mouse Astrocyte TNF-α Viability Assay
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Briefly, mouse astrocytes were treated with TNF-α for 24 h, the cell viability was assayed by cell viability assay kit (Promega, G7570). The luminescent signal recording from the reaction was as the standard of cell viability.
7
Comprehensive Cell Culture Protocol
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RPMI-1640 was bought from Hyclone (Pittsburgh, CA, United States). Foetal bovine serum was purchased from BI (Beit HaEmek, Israel). A cell viability assay kit was purchased from Promega (Madison, Wisconsin, United States). An apoptosis detection kit (PI/Annexin V-FITC) and ROS assay kit (DCFH-DA) were purchased from Genview (Pompano Beach, FL, United States). The reduced glutathione assay kit was purchased from Jiancheng Bioengineering Institute (Nanjing, China). β-ME was purchased from MP Biomedicals (Santa Ana, CA, United States). SASP, NAC, and bovine serum albumin were purchased from Sigma (St Louis, MO, United States). Culture plates/flasks/dishes were acquired from Corning (Tewksbyry, MA, United States). Western blot analysis-related devices and reagents were acquired from Bio-Rad (Hercules, CA, United States). For immunohistochemistry staining, we used the rabbit two-step method kit (Nakasugi Jinqiao, PV-6001). The xCT and GAPDH primary antibody were purchased from Proteintech (Wuhan, Hubei, P.R.C, 26864-1-AP, 10494-1-AP). For the colony formation experiment, we used crystal violet (C0775, Sigma-Aldrich). The medium and lysate that were necessary for organoid culture were acquired from ACCURATE biotechnology.
8
Cell Viability Assay for U87 and LN229 Cells
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We inoculated the treated U87 and LN229 cells in 96-well plates at a density of 1 × 103 cells/well for 24, 48, 72, 96, and 120 h. The plates were examined using a cell viability assay kit (Promega Corp., Fitchburg, WI, United States) in accordance with the manufacturer’s protocol, as described previously (Wang et al., 2021 (link)).
9
Cell Viability Assay for VPA and TMZ
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Cells were seeded and grown overnight and treated with 2-16 mM VPA (Sigma-Aldrich Chemical Co., St Louis, MO), 50 μM of TMZ (Sigma-Aldrich Chemical Co.), or negative control (solvent only) for up to 72 h. At the end of each experiment, cell viability was assessed using the cell viability assay kit (Promega, Madison, WI), according to the manufacturer's instructions. Briefly, 25 µL of treated cell samples were collected to which was added 5 µL of cell viability assay reagent per well. The plates were incubated at 37°C to allow the cells to convert. Then the fluorescent signal was measured with an automatic microplate reader (BioTek Instruments, Winooski, VT). The data were summarized and expressed as percentage of the control.
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