For human HNSCC cell line analysis, cells were seeded at 1x104 cells/well, were left to rest for 24 hours in 37°C and infected with 1, 10, 100 and 1000 VP/cell of Ad5/3-E2F-D24-hTNFa-IRES-hIL-2 or Ad5/3-E2F-D24. Then, the cell viability of the HNSCC cell lines was determined by MTS on day 2, 6, 10. Wells were incubated for 2 hours with 40 µl of CellTiter 96 AQueous One Solution Proliferation Assay reagent (Promega, Wisconsin, USA). Absorbance was read at 492 nm using a Fluostar OPTIMA analyzer (BMG Labtech, Offenburg, Germany). Each time data-point was normalized to the vehicle control group.
Celltiter 96 aqueous one solution proliferation assay reagent
Manufactured by Promega
Sourced in United States
The CellTiter 96 AQueous One Solution Proliferation Assay reagent is a colorimetric method for determining the number of viable cells in proliferation or cytotoxicity assays. The reagent contains a tetrazolium compound that is bioreduced by cells into a colored formazan product, which can be measured spectrophotometrically.
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Lab products found in correlation
6 protocols using celltiter 96 aqueous one solution proliferation assay reagent
1
HNSCC Cell Viability Assay
2
Oncolytic Adenovirus Cytotoxicity Assay
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Human cell lines A549 and RD were plated in 96-well plates (flat bottom) in triplicates at 1 × 104 cells/well for 24 hours and infected with 1, 10, 100 or 1000 VP/cell of either Ad5/3-E2F-d24 virus (also referred in the text as backbone or unarmed backbone), or Ad5/3-E2F-d24-hIL7 (also referred in the text as IL7 virus). Similarly, hamster cell lines HT100, DDT1-MF2 and HapT1 were plated in triplicates at 1 × 104 cells/well for 24 hours and infected with 100, 1000, 5000 or 10000 VP/cell of either backbone virus, or Ad5/3-E2F-d24-hIL7.
Cell viability was measured after 4 days (A549 and RD), 5 days (HT100 and DDT1-MF2) or 8 days (HapT1) by incubating cells for 2 hours with 20% of CellTiter 96 AQueous One Solution Proliferation Assay reagent (Promega, Wisconsin, USA). Absorbance was read at 490 nm using Hidex Sense plate reader (Hidex, Turku, Finland). Data were normalized to the uninfected mock control group.
Cell viability was measured after 4 days (A549 and RD), 5 days (HT100 and DDT1-MF2) or 8 days (HapT1) by incubating cells for 2 hours with 20% of CellTiter 96 AQueous One Solution Proliferation Assay reagent (Promega, Wisconsin, USA). Absorbance was read at 490 nm using Hidex Sense plate reader (Hidex, Turku, Finland). Data were normalized to the uninfected mock control group.
3
Cytotoxicity Assay of CQ, BAF-A1, NH4Cl
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The cytotoxicity of CQ, BAF-A1, and NH4Cl was determined using the CellTiter 96 Aqueous One solution proliferation assay reagent (Promega, Madison, WI, USA). Briefly, these compounds were added at the indicated concentrations to overnight-cultured RD or HEK-293 cells, which were then incubated for the indicated time intervals. Subsequently, 20 µL of the proliferation assay reagent was added to each well of the 96-well plate. The plate was then analyzed at the absorbance of 490 nm after 2 h of incubation at 37 °C.
4
Tumor Cell Viability Assay
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Tumor cells (3 × 105) were plated in triplicate in a 96-well plate and treated with 100 VP/cell of either Ad5/3-E2F-d24-TNFα-IRES-IL-2, aPD-1, or a combination of both. Cell viability was measured on days 3 and 5 by incubating cells for 2 h with 20% of CellTiter 96 AQueous One Solution Proliferation Assay Reagent (Promega, WI, USA). The absorbance was read at 490 nm using a Hidex Sense plate reader (Hidex, Turku, Finland). The data were normalized to those of the uninfected control group.
5
Cell Viability Assay for OVCA Tumor Cultures
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The cell viability of OVCA ex vivo tumor cultures was determined in accordance with a protocol previously described by our group.28 (link) Briefly, cell viability was verified at days 3 and 6 or 7 by incubating 2–3 hours 50 μL of CellTiter 96 AQueous One Solution Proliferation Assay reagent (Promega, Wisconsin, USA). Absorbance was read at 492 nm using a Fluostar OPTIMA analyzer (BMG Labtech, Offenburg, Germany). Each time data point was normalized to the vehicle control group.
6
Cell Viability Assay for Patient Samples
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Cell viability was measured by incubating patient sample for 2 h with 20% of CellTiter 96 AQueous One Solution Proliferation Assay reagent (Promega, Madison, WI, USA). Absorbance was read at 490 nm using a Hidex Sense plate reader (Hidex, Turku, Finland). Data was normalized to the uninfected mock control group.
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