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Alamar blue fluorescent assay

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Alamar Blue is a fluorescent assay used to measure cell viability and proliferation. It utilizes a blue, non-fluorescent dye that is reduced to a red, fluorescent dye by metabolically active cells. The intensity of the fluorescent signal is proportional to the number of viable cells, making it a useful tool for various cell-based assays.

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Lab products found in correlation

Calcusyn, by Biosoft (3 mentions) Xcelligence technology, by Agilent Technologies (2 mentions) E 16 well plates, by Agilent Technologies (2 mentions) Flexstation 3, by Molecular Devices (1 mentions) Flexstation 3 system, by Molecular Devices (1 mentions)

9 protocols using alamar blue fluorescent assay

1

Cell Proliferation Measurement Utilizing AlamarBlue

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Proliferation was assessed using AlamarBlue® fluorescent
assay (Invitrogen, Frederick, MD, USA), at day 4, 14, and 21,
according to the manufacturer’s instructions. We observed a
linear relationship between AlamarBlue fluorescence and the
cell number (data not shown). Fluorescence was measured in
medium samples at 530 nm excitation and 590 nm emission
using a Synergy HT spectrophotometer.
Mokhtari-Jafari F., Amoabediny G., Dehghan M.M., Helder M.N., Zandieh-Doulabi B, & Klein-Nulend J. (2019). Short Pretreatment with Calcitriol Is Far Superior to Continuous Treatment in Stimulating Proliferation and Osteogenic Differentiation of Human Adipose Stem Cells. Cell Journal (Yakhteh), 22(3), 293-301.
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2

Cell Proliferation Quantification in Biomaterials

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Proliferation was assessed by determining cell number in BCP60/40- and BCP20/80-based composites and fibrin gels at days 1 and 11 by using alamarBlue® fluorescent assay (Invitrogen, Frederick, MD, USA), according to the manufacturer's instructions. We found a linear relationship between alamarBlue fluorescence and cell number (data not shown). Fluorescence was read in medium samples at 530 nm with a Synergy HT spectrophotometer.
van Esterik F.A., Zandieh-Doulabi B., Kleverlaan C.J, & Klein-Nulend J. (2016). Enhanced Osteogenic and Vasculogenic Differentiation Potential of Human Adipose Stem Cells on Biphasic Calcium Phosphate Scaffolds in Fibrin Gels. Stem Cells International, 2016, 1934270.
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3

Cell Proliferation Measurement via Alamar Blue

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Cell proliferation of transfected cell lines was measured by the Alamar Blue fluorescent assay (Life Technologies). Briefly, cells were seeded in 96-well plates at a density of 3,000-5,000/well. Basal, 24 h and 48 h cell viability was determined by measurement of fluorescent signal exciting at 560 nm and reading at 590 nm (Flex Station 3; Molecular Devices) at 570 nm. Specifically, the day of measurement, cells were incubated for 3 h in 10% alamar blue/ serum free-media, and then, alamar reduction was measured. Results are expressed as percentage vs. control (mock transfected cells). Medium was replaced by fresh medium immediately after each measurement. In all instances, cells were seeded per quadruplicate and all assays were repeated a minimum of four times.
Sampedro-Núñez M., Luque R.M., Ramos-Levi A.M., Gahete M.D., Serrano-Somavilla A., Villa-Osaba A., Adrados M., Ibáñez-Costa A., Martín-Pérez E., Culler M.D., Marazuela M, & Castaño J.P. (2015). Presence of sst5TMD4, a truncated splice variant of the somatostatin receptor subtype 5, is associated to features of increased aggressiveness in pancreatic neuroendocrine tumors. Oncotarget, 7(6), 6593-6608.
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4

Cell Proliferation and Viability Assays

Cited 1 time
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Cell proliferation was assessed using E-16-well plates and xCELLigence technology (Acea Bioscience, San Diego, CA, USA, distributed by Roche).19 (link) Cell growth was monitored for 72 h. Microelectrodes, placed on the bottom of plates, were used to detect impedance changes proportional to the number of adherent cells and expressed as the cell index. The impedance value of each well was automatically monitored by the xCELLigence system and expressed as a cell index value. The experiments were conducted in triplicate and repeated twice.
For viability assays, melanoma cell lines were plated in 96-well plates at 5000 cells/well in complete medium. 24 hours after plating, varied doses of inhibitors were added in triplicate. 0.1% DMSO was used as negative control. Cell viability was evaluated after 72-hour incubation with drugs using Alamar Blue fluorescent assay (Life Technologies).
Araiza-Olivera D., Feng Y., Semenova G., Prudnikova T.Y., Rhodes J, & Chernoff J. (2017). Suppression of RAC1-driven malignant melanoma by Group A PAK inhibitors. Oncogene, 37(7), 944-952.
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5

Cytotoxicity Evaluation of Compounds

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MEFs were plated in triplicate for each dose of the compound on 96-well plates at 10000 cells/well in complete medium. Twenty-four hours after plating, cells were treated with carrier alone (0.1% DMSO) or bioactive compounds. Cell viability was evaluated seventy-two hours after addition of drugs using alamarBlue fluorescent assay (Life Technologies, USA). The values of IC50, the drug concentration required for 50% growth inhibition, were calculated using software package Calcusyn (Biosoft).
Semenova G., Stepanova D., Deyev S.M, & Chernoff J. (2017). Medium Throughput Biochemical Compound Screening Identifies Novel Agents for Pharmacotherapy of Neurofibromatosis Type I. Biochimie, 135, 1-5.
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6

MPNST Cell Viability Assay

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MPNST cell lines were plated in 96-well plates at 5000 cells/well in complete medium. 24 hours after plating, varied doses of inhibitors were added in triplicate. 0.1% DMSO was used as negative control. Cell viability was evaluated after 72-hour incubation with drugs using alamarBlue fluorescent assay (Life Technologies). Synergistic effects were determined by the Chou/Talalay method (45 (link)). IC50 and CI values were calculated using Calcusyn (Biosoft); (CI) < 1 was defined as synergism. Frax1036 and PD-901 were assumed as having independent modes of action and therefore mutually nonexclusive.
Semenova G., Stepanova D.S., Dubyk C., Handorf E., Deyev S.M., Lazar A.J, & Chernoff J. (2017). Targeting Group I p21-Activated Kinases to Control Malignant Peripheral Nerve Sheath Tumor Growth and Metastasis. Oncogene, 36(38), 5421-5431.
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7

Cell Proliferation and Viability Assays

Cited 1 time
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Cell proliferation was assessed using E-16-well plates and xCELLigence technology (Acea Bioscience, San Diego, CA, USA, distributed by Roche).19 (link) Cell growth was monitored for 72 h. Microelectrodes, placed on the bottom of plates, were used to detect impedance changes proportional to the number of adherent cells and expressed as the cell index. The impedance value of each well was automatically monitored by the xCELLigence system and expressed as a cell index value. The experiments were conducted in triplicate and repeated twice.
For viability assays, melanoma cell lines were plated in 96-well plates at 5000 cells/well in complete medium. 24 hours after plating, varied doses of inhibitors were added in triplicate. 0.1% DMSO was used as negative control. Cell viability was evaluated after 72-hour incubation with drugs using Alamar Blue fluorescent assay (Life Technologies).
Araiza-Olivera D., Feng Y., Semenova G., Prudnikova T.Y., Rhodes J, & Chernoff J. (2017). Suppression of RAC1-driven malignant melanoma by Group A PAK inhibitors. Oncogene, 37(7), 944-952.
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8

MPNST Cell Viability Assay

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MPNST cell lines were plated in 96-well plates at 5000 cells/well in complete medium. 24 hours after plating, varied doses of inhibitors were added in triplicate. 0.1% DMSO was used as negative control. Cell viability was evaluated after 72-hour incubation with drugs using alamarBlue fluorescent assay (Life Technologies). Synergistic effects were determined by the Chou/Talalay method (45 (link)). IC50 and CI values were calculated using Calcusyn (Biosoft); (CI) < 1 was defined as synergism. Frax1036 and PD-901 were assumed as having independent modes of action and therefore mutually nonexclusive.
Semenova G., Stepanova D.S., Dubyk C., Handorf E., Deyev S.M., Lazar A.J, & Chernoff J. (2017). Targeting Group I p21-Activated Kinases to Control Malignant Peripheral Nerve Sheath Tumor Growth and Metastasis. Oncogene, 36(38), 5421-5431.
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9

Cell Proliferation Assay of Ghrelin Variant

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As previously reported [39 (link), 42 (link)], cell proliferation of cell lines transfected with In1-ghrelin or empty (mock) vectors was measured using the alamar-blue fluorescent assay (Life Technologies, Madrid, Spain). Briefly, transfected cells were seeded in 96-well plates at a density of 3000–5000 per well and serum-starved for 12 h. Then, after 3 h of incubation with 10% alamar-blue serum-free medium, basal proliferation rate was obtained by measuring the fluorescent signal exciting at 560 nm and reading at 590 nm using the FlexStation III system (Molecular Devices, Sunnyvale, CA, USA). Subsequently, proliferation rate was similarly measured at 24, 48 and 72 h after the basal proliferation rate evaluation. Medium was replaced by fresh medium containing FBS immediately after each measurement. Results were expressed as percentage referred to control (mock transfected cells). In all experiments, cells were seeded per quadruplicate and all experiments were performed a minimum of four times.
Luque R.M., Sampedro-Nuñez M., Gahete M.D., Ramos-Levi A., Ibáñez-Costa A., Rivero-Cortés E., Serrano-Somavilla A., Adrados M., Culler M.D., Castaño J.P, & Marazuela M. (2015). In1-ghrelin, a splice variant of ghrelin gene, is associated with the evolution and aggressiveness of human neuroendocrine tumors: Evidence from clinical, cellular and molecular parameters. Oncotarget, 6(23), 19619-19633.
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