NIH3T3 empty vector cells (3T3 control cells) for negative control, and NIH3T3 PTPRG-RAF1-expressing cells (3T3 PR cells) (1x103 cells/well) were plated in a 96-well plate and allowed to settle overnight. After 24 h, the medium in each well was replaced with 10 µL of CellTiter 96 AQ One Solution reagent (G3582; Promega, Madison, WI, USA) in 90 µL of DMEM. The plates were incubated at 37°C for 4 h, in a humidified, 5% CO2 atmosphere. Absorbance was recorded at 490 nm using a Spectramax 190 plate reader (Molecular Devices, San Jose, CA, USA). The assay was carried out every 24 h for 96 h in triplicate. A t-test was performed to evaluate significant differences between the two groups.
Celltiter 96 aq one solution reagent
Manufactured by Promega
Sourced in United States
The CellTiter 96® AQ One Solution Reagent is a colorimetric assay for determining the number of viable cells in a sample. The reagent utilizes a tetrazolium compound that is bioreduced by metabolically active cells, creating a colored formazan product that can be measured spectrophotometrically.
Automatically generated - may contain errors
Lab products found in correlation
Spectramax 190, by Molecular Devices (1 mentions)
Glomax multi detection system, by Promega (1 mentions)
Anti ddk antibody, by OriGene (1 mentions)
Pcmv6 entry vector, by OriGene (1 mentions)
Spectramax 96 well plate reader, by Molecular Devices (1 mentions)
Cell cycle detection kit, by Keygen Biotech (1 mentions)
6 protocols using celltiter 96 aq one solution reagent
1
Cell Proliferation Assay of PTPRG-RAF1 Expressing Cells
2
NANOS1 Mutant Effects on TCam-2 Cell Viability
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TCam-2 cells (2 × 105) were transfected with 1.5 µg of constructs encoding the wild-type, the mutated NANOS1 p.[(Pro34Thr);(Ser83del)] or the empty pCMV6-entry vector (OriGene Technologies, PS100001, Rockville, Maryland, USA) in three biological replicates (three independent transfections; five technical repeats in each of five different wells of a 96-well plate) and were cultured for 24, 48, and 72 h after transfection. In order to obtain a similar number per well of viable cells expressing the different constructs for the MTS assay, viable cells were counted 12 h post-transfection using Trypan Blue staining. Similar numbers of cells were seeded in five different wells of a 96-well plate. The cell viability was measured using GloMax®-Multi Detection System (Promega, Madison, Wisconsin, USA) at a 450 nm (and 750 nm for the cell background) wavelength 24, 48, and 72 h after transfection and 4 h after adding 20 µl of CellTiter 96® AQ One Solution Reagent (Promega, Madison, Wisconsin, USA) to each well containing transfected TCam-2 cells. The overexpression of the wild-type and the mutated NANOS1 was measured 12, 24, 48, and 72 h after transfection by western blot using an anti-DDK antibody (OriGene Technologies, TA50011, Rockville, Maryland, USA). VCL (vinculin) protein was used as a loading control.
3
Cell Viability Assay Protocol
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The cell viability assays were assessed using CellTiter 96® AQ One Solution Reagent (Promega, USA) according to the manufacturer's instructions.
4
Cell Viability Assay with siRNA
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HCC2429 or HS-SY-II cells were seeded into 6-well plates, and 50 nM siRNAs were transiently transfected into these cells. Twenty-four hours later, 5,000 cells were plated into 96-well plates. After 48 hours, the medium was replaced with 10 μL of CellTiter 96 AQ One Solution reagent (G3582, Promega, Madison, WI) with 90 μL of DMEM per well. The plate was incubated at 37°C for 4 hours in a humidified atmosphere with 5% CO2, and the absorbance was recorded at 490 nm using a SpectraMax 96-well plate reader (Molecular Devices, San Jose, CA).
5
Cell Viability, Cycle, and Colony Assays
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The cell viability assays were performed using the CellTiter 96® AQ One Solution Reagent (Promega, USA). The cell cycle assays were performed using a Cell Cycle Detection Kit (KeyGEN BioTECH, China). All procedures were performed according to the manufacturer's instructions. For the cell colony formation assay, 1000 cells/well were seeded in a 6-well plate and cultured for 10 days. The colonies were then fixed and stained with a 0.5% crystal violet methanol solution for 30 min and scanned to generate images.
6
Cell Viability, Apoptosis, and Colony Assays
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Cell viability assays were assessed using the CellTiter 96® AQ One Solution Reagent (Promega, USA) according to the manufacturer’s instructions. An apoptosis assay was performed by FACS with Annexin V-FITC/PI staining. For colony formation assays, cells were seeded in 6-well plates at a density of 800 cells/well and cultured with different concentrations of vitamin C at 37 °C for 7 days.
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