Experiments using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide (MTT) methodology were carried out using the Cell Proliferation Reagent (WST-1) assay kit (Roche, Mannheim, Germany) according to the manufacturer's instructions. In brief, for Daoy and D556 cells, 2000 cells per well were seeded in a 96-well plate and incubated with the indicated inhibitors. After 5 days, 10% (v/v) WST-1 reagent was added to each well and absorbance at 450nm was analyzed (using absorbance at 600nm as a reference wavelength), using an Epoch Plate reader and Gen5 software from BioTek Instruments Inc. For proliferation, cells were counted using an automated cell counter (Scepter, Millipore).
Cell proliferation reagent wst 1 assay kit
Manufactured by Roche
Sourced in Switzerland
The Cell Proliferation Reagent (WST-1) assay kit is a colorimetric assay for the quantification of cell proliferation and viability. The kit contains the water-soluble tetrazolium salt WST-1, which is reduced by metabolically active cells to form a colored formazan dye. The absorbance of the formazan dye is directly proportional to the number of viable cells in the sample.
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3 protocols using cell proliferation reagent wst 1 assay kit
1
Cell Proliferation Assay with WST-1
2
Cell Proliferation Assay with WST-1
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Cell proliferation was measured by a Cell Proliferation
Reagent WST-1 Assay Kit (Roche, Basel, Switzerland).
Briefly, siRNAs transfected cells and control
cells were seeded at a concentration of 3×103 cells per
well in 96-well plates (Corning Inc., NY, USA). For
the indicated time, WST-1 solution was applied at
10 μl per well and incubated for 4 hours at 37˚C and
5% CO2. The absorbance [also called optical density
(OD)] was measured with a microplate ELISA reader
(BioTek, Winooski, VT, USA) at 450 nm. Viability
and inhibition rate were calculated according to the
following equations, respectively. Viability (%)=(OD
treated/OD medium)×100%. Inhibition rate (%)=(1-
OD treated/OD control)×100%.
Reagent WST-1 Assay Kit (Roche, Basel, Switzerland).
Briefly, siRNAs transfected cells and control
cells were seeded at a concentration of 3×103 cells per
well in 96-well plates (Corning Inc., NY, USA). For
the indicated time, WST-1 solution was applied at
10 μl per well and incubated for 4 hours at 37˚C and
5% CO2. The absorbance [also called optical density
(OD)] was measured with a microplate ELISA reader
(BioTek, Winooski, VT, USA) at 450 nm. Viability
and inhibition rate were calculated according to the
following equations, respectively. Viability (%)=(OD
treated/OD medium)×100%. Inhibition rate (%)=(1-
OD treated/OD control)×100%.
3
Cell Viability Assay for Alpelisib and MNKi
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