To identify the association between the phosphorylation of CPB1 protein and cell viability, stable cell lines (control, CPB1OE and CPB1OE-mutY) were seeded in 12-well plates at a density of 5×105 cells/well. Cell viability was measured at 6, 24 and 48 h. Cells were collected and mixed with 0.4% trypan blue stain solution (Gibco), after which unstained cells were counted under a microscope (Olympus, Tokyo, Japan).
Trypan blue stain solution
Manufactured by Thermo Fisher Scientific
Sourced in Japan
Trypan blue stain solution is a laboratory reagent commonly used for cell counting and viability assessment. It is a dye that selectively colors dead cells blue, allowing them to be easily distinguished from live cells under a microscope. The solution is typically used in combination with a hemocytometer or other cell counting devices to determine the total number of cells and the percentage of viable cells in a sample.
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Lab products found in correlation
2 protocols using trypan blue stain solution
1
Phosphorylation Impacts Cell Viability
2
Neuronal Viability and Membrane Integrity Assays
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MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay was carried on 96 wells plated neuronal cultures. After each treatment, 20 μl of MTT (5 mg/ml) was added into each well and the plate was further incubated for 1 h at 37 °C. Media in each well was discarded and the colored products of MTT in cells were solubilized with 200 μl of DMSO. Optical intensity at 570 nm was read using Epoch, Biotek multiwell plate reader. Each group contained 6 independent measurements.
Trypan blue stain assay was performed adding 10 μL of Trypan blue stain solution (Gibco) to cultured cells media at the end of rotenone exposure time in order to stain in blue the cytoplasm of cells with damaged plasma membrane. Immediately after the addition of Trypan blue, cells were examined under an inverted microscope (Nikon Eclipse TS100) using a 10× objective (100× magnification) and brightfield images were acquired from three different fields randomly choose per plate per each experimental condition. Then, the number of Trypan blue positive cells per field was accessed.
Trypan blue stain assay was performed adding 10 μL of Trypan blue stain solution (Gibco) to cultured cells media at the end of rotenone exposure time in order to stain in blue the cytoplasm of cells with damaged plasma membrane. Immediately after the addition of Trypan blue, cells were examined under an inverted microscope (Nikon Eclipse TS100) using a 10× objective (100× magnification) and brightfield images were acquired from three different fields randomly choose per plate per each experimental condition. Then, the number of Trypan blue positive cells per field was accessed.
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