Cell viability was determined by the water-soluble tetrazolium (WST)-1 method using the WST-1 cell proliferation and cytotoxicity assay kit (Beyotime, Shanghai, China). Briefly, 5 × 103 cells were seeded in 200 μL/well culture medium in 96-well plates for 24 h and treated with Rap or HA for 72 h. After incubation with WST-1 reagent for 2 h at 37°C, absorbance (450 nm) was measured using an automated microplate reader (Bio-Rad 5 Model 550, Bio-Rad, Hercules, CA, USA). Cell viability percentage = mean optical density (OD) of one experimental group/mean OD of the control × 100%.
5 model 550
Manufactured by Bio-Rad
Sourced in United States
The Bio-Rad Model 550 is a microplate reader designed for absorbance-based assays. It can measure optical density at multiple wavelengths. The device is capable of performing quantitative and qualitative analyses of samples in a 96-well microplate format.
Automatically generated - may contain errors
3 protocols using 5 model 550
1
WST-1 Cell Viability Assay
2
Cell Viability and Colony Formation Assays
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Water soluble tetrazolium (WST)-1 assay was performed to measure cell viability after transfection. 5×103 of transfected cells per well were seeded into 96-well plates and cultured for 24, 48, 72, or 96 h. Afterward, cells were then incubated with WST-1 reagent for 2 h at 37°C. The absorbance at 450 nm was measured with the use of an automated microplate reader (Bio-Rad 5 Model 550, Bio-Rad, Hercules, CA, USA) and the growth curve was calculated.
For colony formation assay, 1×103 of transfected cells per well were seeded in 6-well plates and grown in complete media at 37°C. After 14 days for culture, colonies were stained with crystal violet and the number of colonies was counted by automated colony counter (Alpha Innotech, San Leandro, CA, USA).
For colony formation assay, 1×103 of transfected cells per well were seeded in 6-well plates and grown in complete media at 37°C. After 14 days for culture, colonies were stained with crystal violet and the number of colonies was counted by automated colony counter (Alpha Innotech, San Leandro, CA, USA).
3
Cytotoxicity Evaluation of Graphene Oxide
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The cells were seeded in 96-well flat-bottomed microtiter plates (5 × 10 3 cells in 200 µL per well) for 36 h and treated with different concentrations of GO, GO-BENA and distilled water (Laboratory homemade). The percentage of living cells was determined using the Lactate Dehydrogenase (LDH) Cytotoxicity Assay Kit and WST-1 Cell proliferation and cytotoxicity assay kit (Beyotime Institute of Biotechnology, Jiangsu, China). Assays were performed in accordance with the manufacturer's instructions. The absorbance at 450 nm was monitored based on the absorbance maximum of the formazan dye and the reference wavelength was set at 630 nm. The data was measured using an automated microplate reader (Bio-Rad 5 Model 550, Bio-Rad, Hercules, CA, USA). Cytotoxicity percentage(LDH kit) was determined by the equation: Cytotoxicity percentage(%)=(Absorbance of treated samples -Absorbance of control group)/(Maximum enzyme activity absorbance -Absorbance of control group). Cell viability percentage (WST-1 kit) was determined by the mean optical density (OD) of one experimental group divided by the mean OD of the untreated group × 100%.
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