Proliferation was assessed using the colorimetric CellTiter96® Aqueous Non-Radioactive Cell Proliferation Assay (Promega, Madison, WI). Cells (5 × 103) were plated and treated the following day with UAB30 or 6-Me in increasing concentrations (0 to 100 μM), or an equivalent concentration of vehicle, dimethyl sulfoxide (DMSO). CellTiter96® dye (10 μL, Promega) was added 72 hours later and absorbance read at 490nm on a microplate reader (Epoch Microplate Spectrophotometer, BioTek Instruments, Winooski, VT). To assess viability, alamarBlue® colorimetric assay (Thermo Fisher Scientific Inc.) was utilized. Cells (1.5 × 104) were plated on 96-well plates and treated the following day with vehicle, UAB30, or 6-Me in increasing concentrations. After 72 hours, 10 μL of alamarBlue® dye (Thermo Fisher Scientific Inc.) was added and absorbance was read at 570nm and 600nm. Results of proliferation and viability from at least three biologic replicates were reported as fold change ± standard error of the mean (SEM).
Celltiter96 dye
Manufactured by Promega
The CellTiter96® dye is a colorimetric assay used to determine the number of viable cells in a given sample. It measures the metabolic activity of cells, providing an indication of their overall health and proliferation.
Automatically generated - may contain errors
Lab products found in correlation
Alamar blue dye, by Thermo Fisher Scientific (3 mentions)
Epoch microplate spectrophotometer, by Agilent Technologies (3 mentions)
Celltiter 96 aqueous non radioactive cell proliferation assay, by Promega (2 mentions)
Alamarblue colorimetric assay, by Thermo Fisher Scientific (2 mentions)
Biotek gen5, by Agilent Technologies (1 mentions)
Celltiter 96 aqueous one solution cell proliferation assay, by Promega (1 mentions)
4 protocols using celltiter96 dye
1
Cellular Proliferation and Viability Assay
2
Assessing Tumor Cell Viability and Proliferation
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Following tumor dissociation, cells were suspended in NB media and counted. To assess cell survival, alamarBlue® colorimetric assay (Thermo Fisher Scientific) was utilized. Cells (1.5 × 104) were plated on 96-well plates and treated the following day with PF or Y15 in increasing concentrations, or an equivalent concentration of vehicle (DMSO). After 24 hours, 10 μL of alamarBlue® dye (Thermo Fisher Scientific) was added and absorbance was read at 570 nm and 600 nm on a microplate reader (Epoch Microplate Spectrophotometer, BioTek Instruments, Winooski, VT). Proliferation was evaluated using the colorimetric CellTiter96® Aqueous Non-Radioactive Cell Proliferation Assay (Promega, Madison, WI). Cells (5 × 103) were plated and treated the following day with vehicle, PF, or Y15 in increasing concentrations. CellTiter96® dye (10 μL, Promega) was added 24 hours later and absorbance read at 490 nm. Results in survival and proliferation from at least three biologic replicates were reported as fold change ± SEM.
3
Evaluating Neuroblastoma Cell Proliferation
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The CellTiter 96® assay was used to assess the effect of RA and UAB30 on proliferation in neuroblastoma PDX cells in vitro. Both COA3 and COA6 cells (1 × 104 cells/well) were plated in a 96-well plate, and treated with increasing doses of RA or UAB30 (0, 10, 25, 50, 100 μM) for 96 h. CellTiter 96® dye (10 μL, Promega, Madison, WI) was then added to each well and the absorbance was measured at 490 nm using a microplate reader (BioTek Gen5, BioTek Instruments, Winooski, VT). Similarly, COA3 or COA6 cells (3 × 104 cells/well) were plated in a 96-well plate to perform the alamarBlue® assay to measure viability. Cells were treated with increasing doses of RA and UAB30 (0, 10, 25, 50, 100 μM) for 96 h. AlamarBlue® dye (10 μL, Thermo Fisher Scientific, Waltham, MA) was added to each well and the absorbance at 562 nm (reduced reagent) and 595 nm (oxidized reagent) was measured using a microplate reader (BioTek Gen5).
4
Proliferation Assay for Cell Lines
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Proliferation was examined using the CellTiter 96® Aqueous One Solution Cell Proliferation assay (Promega). Cells (5 × 103 cells) were plated onto 96-well plates. After 24 h, CellTiter 96® dye (10 µL) was added to each well and the absorbance was measured at 490 nm using a microplate reader (Epoch Microplate Spectrophotometer, BioTek Instruments, Winooski, VT, USA). For the siRNA experiments, we transfected the cells with siRNA for 72 h as described above, then we lifted and plated transfected cells onto 96-well plates for 24 h, added CellTiter96® dye (Promega), and read the plates. Proliferation experiments were completed with at least three biologic replicates and data reported as fold change ± standard error of the mean (SEM).
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