Cell proliferation was measured by WST-1 assay kit (Roche, Basel, Switzerland). Briefly, siRNAs transfected cells and control cells were seeded at a concentration of 3 × 103 cells per well in 96-well plates (Corning Inc., NY, USA). For indicated time, WST-1 solution was applied at 10 μl per well and incubated for 4 hours at 37°C, 5% CO2. The absorbance was measured with a microplate ELISA reader (BioTek, Winooski, VT, USA) at 450 nm.
Wst 1 assay kit
Manufactured by Roche
Sourced in Germany, Switzerland, United States
The WST-1 assay kit is a colorimetric method for the non-radioactive quantification of cell proliferation, viability, and cytotoxicity. The assay utilizes the tetrazolium salt WST-1, which is cleaved to a colored formazan dye by cellular enzymes. The amount of formazan produced is directly proportional to the number of metabolically active cells in the culture, and can be measured using a spectrophotometer.
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Lab products found in correlation
Varioskan flash, by Thermo Fisher Scientific (4 mentions)
Carbachol, by Abcam (2 mentions)
Donepezil, by Abcam (2 mentions)
1 1 dimethyl 4 diphenylacetoxypiperidinium iodide 4 damp, by Abcam (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Multiskan fc, by Thermo Fisher Scientific (2 mentions)
Rpmi 1640, by Thermo Fisher Scientific (2 mentions)
Elisa microplate reader, by Agilent Technologies (1 mentions)
96 well plate, by Corning (1 mentions)
Multimode plate reader, by Agilent Technologies (1 mentions)
Ldh assay kit, by Roche (1 mentions)
Benchmark microplate reader, by Bio-Rad (1 mentions)
Cyquant assay, by Thermo Fisher Scientific (1 mentions)
Prism 5, by GraphPad (1 mentions)
Rpmi 1640, by Lonza (1 mentions)
Penicillin, by Lonza (1 mentions)
Streptomycin, by Lonza (1 mentions)
L glutamine, by Lonza (1 mentions)
Varioskan, by Thermo Fisher Scientific (1 mentions)
58 protocols using wst 1 assay kit
1
Cell Proliferation Assay using WST-1
2
Evaluating Cancer Cell Proliferation with WST-1 Assay
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A water-soluble tetrazolium-1 (WST-1) assay kit (Roche Diagnostics GmbH, Germany) was used to determine the proliferative potential of cancer cells. Cells were seeded and cultured at 1 × 104 cells per well in 96-well plates for 24 h and then treated with EPA or DHA solutions for 24 and 48 h or FFAE of krill oil for 24 h. All treatments were performed in triplicates and the concentrations of EPA, DHA and Oxaliplatin were selected based on their respective dose-response curves. Four concentrations (50 μM, 100 μM, 200 μM and 250 μM) of DHA and three concentrations (50 μM, 100 μM, 200 μM) of EPA were used. FFAE of KO were diluted in ethanol at three concentrations: 0.03 μL, 0.06 μL and 0.12 μL/100 μL prior to the treatment and that equate to the concentrations of EPA and DHA per 100 μL well at 0.13/0.06, 0.26/0.13 and 0.52/0.26 μM, respectively. In all experiments, 0.1% ethanol was used as a vehicle control, non-treated cells as a negative control, and Oxaliplatin as a positive control.. The WST-1 reagent (10 μL) was added to each well after respective treatment time point and incubated at 37 °C for 1 h. Cell proliferation was measured using a micro-plate reader (Varioskan Flash, Thermo Scientific) at the absorbance of 450 nm. Each experiment was repeated three times for each cell line.
3
Cell Proliferation and Colony Formation
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For cell proliferation detection, the cancer cells (1000 cells/well) were seeded in 96-well plates and then detected using a WST-1 Assay Kit (Roche) for succession days. For colony formation, the cancer cells (500 cells/well) were seeded into the 6-well plates for 2 weeks, and then 1% crystal violet solution was used to stain all cell colonies. Finally, we take pictures for colonies and count the numbers of colonies.
4
Cell Proliferation Assay Protocol
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Cells were seeded in 96 well plates (1 × 104 cells per well) 24 h prior to treatments. To assess cell proliferation, cells were assessed after 48 h using a WST-1 assay kit (Roche, Mannheim, Germany) as per manufacturer’s instructions. Absorbance was measured at a wavelength of 450 nm on a microplate reader (Bio-Rad Laboratories, Hercule, CA, USA). IC50 was calculated according to standard guidelines.
5
Cell Adhesion Assay Using AT2 Cells
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For cell adhesion assay, the AT2 cells were collected immediately following the corresponding treatments in each group and seeded in fibronectin pre-coated 96-well plates (50,000 cells/well), and cultured for 2 h at 37°C in DMEM supplemented with FBS (Gibco; Thermo Fisher Scientific, Inc.). Subsequently, the plates were washed with PBS buffer to remove any unattached cells, and subsequently blocked with 1% BSA for 30 min at 37°C. The cells were stained for 1 h at 37°C using a WST-1 assay kit (Roche Diagnostics) according to the manufacturer's protocol. The absorbance was measured at 690 nm using a multimode plate reader (BioTek Instruments, Inc.).
6
Quantifying Genistein Cytotoxicity in NHDF and KEL FIB Cells
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The cytotoxic effects of genistein were quantified using a WST-1 assay kit (Roche Diagnostics GmbH, Mannheim, Germany). NHDF and KEL FIB cells were seeded at a density of 1 × 104 cells/well in 96-well plates, before incubating at 37 °C, 5% CO2, for 24 h to facilitate cellular attachment. The medium was then changed and supplemented with genistein to a final concentration of 370, 37, 3.7, 0.37, or 0 µM (control). Cells were cultured in the genistein-containing medium for a further 72 h at 37 °C, 5% CO2, before the medium in each well was replaced by 100 µL of fresh medium plus 10 µL of WST-1 reagent, and the cells were incubated for a further 45 min. During this incubation the stable tetrazolinum salt in the WST-1 reagent is converted to a water soluble formazan dye by any viable cells that are present, in a process that is largely dependent on the bio-reduction of the NAD(P)H produced by glycolysis. This assay thus measures the number of viable cells directly. After completion of the WST-1 assay incubation period, the optical density at 450 nm of each well was measured using a UMV340 microplate reader (Biogenet Asys Hitech GmbH, Austria). The cytotoxicity of all genistein concentrations was assayed in triplicate in three independent experiments.
7
Assessing Viability of Mouse Lung Slices
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Viability of mouse lung slices was tested by measuring leakage of LDH from the lung slices using an LDH assay kit (Roche Applied Science) and enzymatic activity based on the cellular cleavage of water-soluble tetrazolium salt (WST-1) to formazan in the lung slices using a WST-1 assay kit (Roche Applied Science) for up to 6 days in culture. Released LDH and soluble formazan in supernatant of mouse lung slice culture medium were quantified by a spectrophotometer (SpectraMax Plus 384). Mouse lung slices exposed to 0.3% Triton X-100 for 15 min at 37°C served as a positive control.
8
Evaluating Cancer Cell Proliferation via WST-1 Assay
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A water-soluble tetrazolium-1 (WST-1) assay kit (Roche Diagnostics GmbH, Germany) was used to determine the proliferative potential of cancer cells. Cells were seeded and cultured at 1 × 104 cells per well in 96-well plates for 24 h, and then treated with individual PUFA solutions for 24, 48 or 72 h; or free fatty acid extract solutions of oils (krill oil or fish oil) for 48 h. All treatments were performed in quadruplicates. For PUFA treatments, three concentrations of each fatty acid were used including 50 μM, 100 μM and 200 μM. 0.1 % ethanol was used as a vehicle control. Additional assays were performed to observe the effects of various EPA concentrations (100 μM, 120 μM, 140 μM, 160 μM, 180 μM and 200 μM) on cell proliferation of HCT-15 cells after 48 h of treatment. The treatment concentrations of free fatty acid extract of oils (krill oil or fish oil) are 0.03, 0.06, 0.12 or 0.24 μL/100 μL well. 1 % DMSO was used as a vehicle control. Non -treated cells were used as a negative control in all experiments. 10 μL WST-1 reagent was added to each well after respective treatment time points and incubated at 37 °C for one hour. Cell proliferation was measured using a microplate reader (Varioskan Flash, Thermo Scientific) at the absorbance of 450 nm.
9
Proliferation Assay of Mutant KRAS
10
Cell Growth and Clonogenicity Assay
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For cell growth assay, cells (2×103 cells per well) were treated with miR-150-NF or blank PLGA nanoparticles (0.05 mg each), as described above, and cultured for 5 days. Cell growth was then monitored using WST-1 assay kit (Hoffman-La Roche Ltd., Basel, Switzerland) as per manufacturer’s instructions. The absorbance was measured at a wavelength of 450 nm using a Bio-Rad Benchmark microplate reader (Bio-Rad Laboratories Inc.). For clonogenicity assay, cells were cultured in six-well plates until they reached 60%–70% confluence, and were subsequently treated with miR-150-NF or blank PLGA nanoparticles (0.05 mg each). Following 48 hours transfection, cells were trypsinized and plated in six-well plates at a density of 1×103 cells per well in a regular media for colony formation. After 2 weeks, colonies were fixed with methanol, stained with crystal violet, photographed, and counted using image analysis software (Gene Tools; Syngene, Frederick, MD, USA).
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