WST-1 cell proliferation assay (Clontech) and the Dual-Luciferase Reporter Assay System (Promega) was performed according to the manufacturer’s instruction. For colony formation assay, 2000–5000 cells per well were seeded on 6-well plate. After 2–3 weeks, the cells were first fixed by 4% paraformaldehyde and then stained with 0.05% crystal violet. The colonies were then imaged and counted for quantification.
Wst 1 cell proliferation assay
Manufactured by Takara Bio
Sourced in United States, Japan
The WST-1 cell proliferation assay is a colorimetric method for the quantitative determination of cell proliferation and viability. The assay is based on the cleavage of the tetrazolium salt WST-1 to a colored formazan product by cellular mitochondrial dehydrogenases. The amount of the formazan dye formed directly correlates with the number of metabolically active cells in the culture.
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Lab products found in correlation
Dual luciferase reporter assay system, by Promega (1 mentions)
Luciferase assay system, by Promega (1 mentions)
Multiskan plate reader, by Thermo Fisher Scientific (1 mentions)
Protein assay dye reagent concentrate, by Bio-Rad (1 mentions)
Glomax discover, by Promega (1 mentions)
Fluostar omega microplate reader, by BMG Labtech (1 mentions)
9 protocols using wst 1 cell proliferation assay
1
Cell Proliferation and Colony Formation Assay
2
Angpt2 Modulates MMC Viability
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The viability of MMCs was determined by using WST-1 Cell Proliferation Assay (Clontech Laboratories, Mountain View, CA, USA). MMCs (5 × 103 cells/well) were seeded in 96-well plates for 24 hr. Cells were cultured under NG or HG conditions, with or without mouse recombinant Angpt2 protein (300 ng/mL), for 48 hr. Cells were incubated with WST-1 substrate at 37°C for 1 hr, and absorbance at 450 nm was detected using an ELISA reader.
3
Evaluating PI3K/mTOR Inhibitors on PIK3CA Mutant Cells
4
Evaluating Reagents for JEV RNA Replication
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The Firefly luciferase (FL) assay was performed to evaluate the effects of each reagent on JEV RNA replication as described previously [7] (link). Briefly, BHKMuarPACLuc rep cells were plated onto 24-well plates (1 × 105 cells per well) in triplicate and then treated with each reagent at several concentrations for 72 h. After treatment, the cells were subjected to the Luciferase assay system (Promega, Madison, WI) according to the manufacturer's protocol. The 50% effective concentration (EC50) of each reagent was determined from the assay results.
The WST-1 cell proliferation assay was performed to evaluate the cell toxicity of each reagent as described previously [12] (link). Briefly, BHKMuarPACLuc rep cells were plated onto 96-well plates (5 × 103 cells per well) in triplicate and then treated with each reagent at several concentrations for 72 h. After treatment, the cells were subjected to the WST-1 cell proliferation assay (Takara Bio, Otsu, Japan) according to the manufacturer's protocol. The 50% cytotoxic concentration (CC50) of each reagent was determined from the assay results.
The WST-1 cell proliferation assay was performed to evaluate the cell toxicity of each reagent as described previously [12] (link). Briefly, BHKMuarPACLuc rep cells were plated onto 96-well plates (5 × 103 cells per well) in triplicate and then treated with each reagent at several concentrations for 72 h. After treatment, the cells were subjected to the WST-1 cell proliferation assay (Takara Bio, Otsu, Japan) according to the manufacturer's protocol. The 50% cytotoxic concentration (CC50) of each reagent was determined from the assay results.
5
Quantifying Cell Viability via WST-1 Assay
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Cell viability assay was performed using pre-mixed WST-1 cell proliferation assay (Takara). WST-1 reagent contains a tetrazolium salt that is cleaved by mitochondrial reductase enzymes of a viable cell to produce yellow to orange colored formazan dye. Formazan is known to show an absorbance at 440 nm. The intensity of formazan dye depends on the number of viable cells or metabolically active cells. After each treatment period, 10 μL of WST-1 reagent was added to each well and plates were incubated at 37 °C at 5 % CO2 for 2 h. Absorbance was measured in a Multiskan plate reader (Thermo Scientific) at 440 nm. Percent decrease in cell viability was calculated from the absorbance of negative control or untreated cells as per the kit protocol. A two-sample t-test was performed using the function in Microsoft Excel to determine the statistical significance of the data at p < 0.05 and p < 0.01.
6
Cell Viability and Protein Assay
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Cell viability was examined using a WST-1 cell proliferation assay (Takara) according to the manufacturer’s instructions. For amido black staining, cells were washed with PBS and fixed with methanol. Then, 0.5% Amido Black solution was added and cells were incubated for 20 min at room temperature. After 20 min, the solution was removed and stained cells were quantified using EPSON GT-X820. Protein assay was by using Protein Assay Dye Reagent concentrate (Bio-Rad) according to the manufacturer’s instructions.
7
WST-1 Cell Proliferation Assay
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Cell proliferation was evaluated by WST-1 cell proliferation assay (Takara) following the manufacturer’s instructions. Shortly HeLa “scrambled” and EFR3A KnD cells were seeded in 96-well plates at a density of 5000 cells per well and allowed to grow for 24 h. After adding the WST-1 cell proliferation assay reagent, cells were then incubated for 1 h at 37 °C and 5% CO2. Subsequently, absorbance was determined in a GloMax Discover microplate reader (Promega, Southampton, United Kingdom) at 450 nm.
8
Evaluating Cancer Cell Proliferation
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Logarithmically growing cancer cells were seeded at optimized densities (8 × 102 cells/well) for each examined cell line onto individual wells of 96-well polystyrene tissue culture-treated microplates (3598, Corning, Corning, NY). Cells were treated with IRX4647 or vehicle (dimethyl sulfoxide, DMSO) at desired concentrations 24 h later. The WST-1 cell proliferation assay (Takara Bio USA, Inc., San Jose, CA) measured cell growth over 5 days using a FLUOstar Omega microplate reader (BMG Labtech, Ortenberg, Germany). These assays were independently repeated at least three times with each biological replicate performed at least in triplicate. Data were normalized to baseline (0 h; no treatment) cell growth.
9
DNR Cytotoxicity in NKNT-3/NTCP cells
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NKNT-3/NTCP cells were plated onto 96-well plates (0.8 Â 10 4 cells per well) in duplicate and then treated with 0, 0.1, 0.5, or 1.0 mM of DNR for 12 h. At 0, 12, 24, and 48 h after treatment, we performed a WST-1 cell proliferation assay according to the manufacturer's protocol (Takara Bio, Kusatsu, Japan).
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