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Celltiter blue cell viability assay

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The CellTiter-Blue Cell Viability Assay is a fluorometric method used to quantify the number of viable cells in a culture. The assay relies on the ability of living cells to convert a redox dye (resazurin) into a fluorescent end product (resorufin). The amount of fluorescent resorufin produced is proportional to the number of viable cells present in the sample.

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Lab products found in correlation

Caspase glo 3 7 assay, by Promega (10 mentions) Spectramax m5, by Molecular Devices (9 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (8 mentions) Celltiter glo luminescent cell viability assay, by Promega (7 mentions) Glomax multi detection system, by Promega (7 mentions) Microtest 96, by BD (7 mentions) Infinite m200, by Tecan (7 mentions) Facscalibur, by BD (7 mentions) Trypsin edta, by Thermo Fisher Scientific (7 mentions) Celltiter blue reagent, by Promega (6 mentions) Dimethyl sulfoxide (dmso), by Merck Group (6 mentions) Prism 6, by GraphPad (6 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (6 mentions) Apo one homogeneous caspase 3 7 assay, by Promega (5 mentions) Fluostar optima microplate reader, by BMG Labtech (5 mentions) Ovcar3, by American Type Culture Collection (5 mentions) Mda mb 231, by American Type Culture Collection (5 mentions) Glutamax, by Thermo Fisher Scientific (5 mentions) Spectramax m2, by Molecular Devices (4 mentions) Spectramax m2 plate reader, by Molecular Devices (4 mentions)

Close spelling variants of this product

CellTiter-Blue (464 citations) CellTiter-Blue assay (204 citations) CellTiter-Blue viability assay (68 citations)

435 protocols using celltiter blue cell viability assay

1

Anticancer Potential of Crude Extracts

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The anticancer and cytotoxicity of the crude extracts (at a concentration of 100 µg/mL) were assessed using the CellTiterBlue Cell Viability Assay (Promega, Mannheim, Germany) against A-549 lung carcinoma cells, MDA-MB-231 breast cancer cells and HaCaT human keratinocytes (Cell Line Service, Eppelheim, Germany). The cells were seeded into 96-well microplates at a concentration of 1 × 104 cells /well. They were cultivated in RPMI medium augmented with 10% fetal bovine serum, 100 µg/mL penicillin, and 100 mg/mL streptomycin at 37 °C under a humidified atmosphere and 5% CO2.
After 24 hrs of cultivation, the medium was replaced with fresh medium (100 µl) containing the crude extracts, and again cultivated for 24 hrs. Further tests were done according to the CellTiterBlue Cell Viability Assay protocol described by the manufacturer (Promega, Mannheim, Germany). The anticancer drug tamoxifen was used as the positive control. The growth medium and 0.5% DMSO were tested as negative controls. The inhibition rates were computed from fluorescence measurements taken with the microplate reader, with excitation and emission wavelengths of 560 nm and 590 nm, respectively.
Oppong-Danquah E., Passaretti C., Chianese O., Blümel M, & Tasdemir D. (2020). Mining the Metabolome and the Agricultural and Pharmaceutical Potential of Sea Foam-Derived Fungi. Marine Drugs, 18(2), 128.
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2

Cell Viability Assay with ABT199

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The viability of the cells exposed to ABT199 was assessed using the CellTiter-Blue cell viability assay (Promega, Madison, WI, USA, G8081). The cells were seeded in a 96-well plate at a density of 1 × 10⁴ cells per well, and this was carried out in quadruplicate. ABT199 was subsequently added to each of the wells to complete a total volume of 100 µL/well. The plate was incubated at 37 °C for 48 h, followed by the addition of 20 µL of the cell titer-blue reagent to each well and further incubation for 2 h. Fluorescence was measured at 560/590 nm using a multi-mode microplate reader (Spectramax i3, Molecular Devices, LLC, San Jose, CA, USA).
Alkhatabi H.A., Zohny S.F., Shait Mohammed M.R., Choudhry H., Rehan M., Ahmad A., Ahmed F, & Khan M.I. (2022). Venetoclax-Resistant MV4-11 Leukemic Cells Activate PI3K/AKT Pathway for Metabolic Reprogramming and Redox Adaptation for Survival. Antioxidants, 11(3), 461.
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3

Cell Proliferation and Anchorage-Independent Growth

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Cell proliferation in culture was measured using the CellTiter Blue Cell Viability Assay (Promega) according to manufacturer instructions. Anchorage independent growth assays were done by plating a base layer of 1:1 ratio 1% agar and McCoy’s 5A (10%FBS) then a top layer 1:1 ratio 0.6% agar and McCoy’s 5A (10%FBS) with cells seeded at the desired density. Colonies were grown at 37C, 5% CO2 and replenished with media twice per week to avoid dehydration.
Che Y., Siprashvili Z., Kovalski J.R., Jiang T., Wozniak G., Elcavage L, & Khavari P.A. (2019). KRAS regulation by small non-coding RNAs and SNARE proteins. Nature Communications, 10, 5118.
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4

Cell Viability Assay for Titanium Sheets

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Cell viability were determined by a CellTiter-BlueCell Viability Assay (Promega, Madison, WI, USA, G808A), which was used according to the manufacturer’s instructions. After 1, 3, and 7 days, 50 μL of CellTiter-Blue dye was added to coated and non-coated titanium sheets for every 500 μL of culture media, and samples were incubated for 4 h at 37 °C and 5% CO2. Sample fluorescence was analyzed using a spectrophotometer (SmartSpec Plus, Bio-Rad, Hercules, CA, USA) at a wavelength of 600 nm, which generated density optic values.
Camargo S.E., Xia X., Fares C., Ren F., Hsu S.M., Budei D., Aravindraja C., Kesavalu L, & Esquivel-Upshaw J.F. (2021). Nanostructured Surfaces to Promote Osteoblast Proliferation and Minimize Bacterial Adhesion on Titanium. Materials, 14(16), 4357.
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5

HUVEC Adhesion and Proliferation on Collagen-Coated Alginate Microspheres

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HUVECs adhesion and proliferation on collagen-coated alginate microspheres were determined by CellTiter-Blue Cell Viability Assay (Promega, Fitchburg, WI) according to manufacturer’s instructions. Briefly, 4 h, 48 h, and 72 h after cell seeding, the alginate/collagen-HUVECs microspheres were transferred into a new well. 200 μl of assay reagent and 1 mL of culture medium were added. After 4 h incubation, the supernatant was read by fluorescence with excitation 560 nm and emission 590 nm filter pair (SpectraMax M2, Molecular Device, Sunnyvale, CA). Alginate/collagen microspheres without cells were subjected to the same process and data were used as the blank. All the data were normalized to cell number by the standard curve. Three samples were tested for each group.
Yao R., Alkhawtani A.Y., Chen R., Luan J, & Xu M. (2019). Rapid and efficient in vivo angiogenesis directed by electro-assisted bioprinting of alginate/collagen microspheres with human umbilical vein endothelial cell coating layer. International Journal of Bioprinting, 5(2.1), 194.
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6

High-Throughput Drug Screening Protocol

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A panel of 120 drugs, either FDA-approved or in clinical trial was obtained from Selleckchem, (Houston, TX). All drugs in this study were dissolved in 100% dimethylsulfoxide (DMSO, Sigma-Aldrich) and stored as 10 mM stock. For the preliminary drug screen, cells were plated with culture media in 96-well clear bottom black microplates (Corning Costar) at a density of 5 × 103 cells per well and cultured overnight at 37° C with 5% CO2. The next day, cells were treated with either vehicle (DMSO) or one three drug concentrations (0.1 μM, 1 μM, and 10 μM in 0.5% DMSO) in triplicate for 72 h. The drugs were given to the cells at a final concentration of 0.5% DMSO. Cell viability was measured using the CellTiter-Blue Cell Viability Assay (Promega) according to the manufacturer’s instructions and an Infinite M200 Pro microplate reader (Tecan). The potency (IC50) of each drug was calculated by nonlinear least-squares curve-fitting using Prism 6 (GraphPad). Selected drugs were further assessed using a 13-point concentration range with 2-fold dilutions in triplicate to confirm key findings from the drug screen.
Zhang L., Nesvick C.L., Day C.A., Choi J., Lu V.M., Peterson T., Power E.A., Anderson J.B., Hamdan F.H., Decker P.A., Simons R., Welby J.P., Siada R., Ge J., Kaptzan T., Johnsen S.A., Hinchcliffe E.H, & Daniels D.J. (2022). STAT3 is a biologically relevant therapeutic target in H3K27M-mutant diffuse midline glioma. Neuro-Oncology, 24(10), 1700-1711.
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7

Cytokine-Dependent Cell Viability Assay

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To remove the cytokines, Ba/F3-gp130 cell lines were washed three times with sterile PBS. 5 × 103 cells were suspended in DMEM supplemented with 10% FCS, 60 mg/l penicillin, and 100 mg/l streptomycin and cultured for 3 days in a final volume of 100 μl with or without cytokines (applied as conditioned media from transfected CHO-K1 cells) as indicated. The CellTiter-Blue Cell Viability Assay (Promega) was used to estimate viable cells by recording the fluorescence (excitation 560 nm, emission 590 nm) using the Infinite M200 PRO plate reader (Tecan) immediately after adding 20 μl of reagent per well (time point 0) and up to 2 h after incubation under standard cell culture conditions. The fluorescent signal from the CellTiter-Blue Reagent is proportional to the number of viable cells. All values were measured in triplicates per experiment. Fluorescence values were normalized by subtraction of time point 0 values. All experiments were performed at least two times, and one representative experiment was selected. Data are presented as means ± SD. For multiple comparisons, one-way ANOVA, followed by Bonferroni correction, was used (GraphPad Prism 6.0, GraphPad Software Inc). Statistical significance was set at the level of p ≤ 0.05 (∗p ≤ 0.05, ∗∗p ≤ 0.01, ∗∗∗p ≤ 0.001).
Georgy J., Arlt Y., Moll J.M., Ouzin M., Weitz H.T., Gremer L., Willbold D., Grötzinger J., Thives-Kurenbach F., Scheller J, & Floss D.M. (2021). Tryptophan (W) at position 37 of murine IL-12/IL-23 p40 is mandatory for binding to IL-12Rβ1 and subsequent signal transduction. The Journal of Biological Chemistry, 297(5), 101295.
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8

Optimizing UV Exposure for Cell Viability

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Optimal UV exposure time was determined using CellTiter-Blue® cell viability assay (Promega Corporation, USA). Cells were grown to 70–80% confluency in 100 mm culture plates and synchronized by overnight serum starvation. Viability assay was performed by adding dye in 1:5 ratio to the culture medium. Cells were incubated in the dark for ~4 hours and absorbance was recorded using UV-2401 PC spectrometer (Shimadzu). 570 and 600 nm were chosen as reference wavelengths, and blank samples consisted of only CellTiter-Blue reagent without cells. Culture medium background at 600 nm was subtracted from experimental values at 570 nm and plot of this difference against the UVR exposure time was generated. Three different time points where cells did not have any structural damage but had significant differences in the absorbance value of the reduced dye product were chosen for further experiments.
Singh S.P., Kang S., Kang J.W., So P.T., Dasari R.R., Yaqoob Z, & Barman I. (2017). Label-free characterization of ultra violet-radiation-induced changes in skin fibroblasts with Raman spectroscopy and quantitative phase microscopy. Scientific Reports, 7, 10829.
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9

Cytotoxicity of PTX and o(LA)8-PTX Formulations

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The cytotoxicity of PTX and o(LA)8-PTX (free and micelle) against a 4T1-luc breast carcinoma cell lines was investigated by the CellTiter-Blue® Cell Viability Assay (Promega, WI). The cells were seeded into a 96-well plate at a seeding density of 4,000 cells/100 μL/well and cultured in Dulbecco’s Modified Eagle Medium (DMEM) at 37 °C in 5% CO2 incubator for 24 h. PTX, o(LA)2-PTX or o(LA)8-PTX was dissolved in DMSO. The final level of DMSO in the medium was < 0.3%. Each was added into the wells to attain levels of 0.001 – 10 μM. While PTX-PM or o(LA)8-PTX-PM in PBS solution (10 mM, pH 7.4) was added into the wells to attain levels of 0.01 – 100 μM. Cells cultured with diluted DMSO or PBS in medium were used as controls. Treated cells were placed in an incubator at 5% CO2 at 37 °C for 72 h. The medium in each well was aspirated, and 100 μL of 20% (v/v) CellTiter-Blue® reagent in serum free DMEM was added, followed by incubation at 37 °C in 5% CO2 atmosphere for 1.5 h. Fluorescence intensity of viable cells was measured by a SpectraMax M2 plate reader (Molecular Devices, CA) with excitation and emission at 560 and 590 nm, respectively. The half maximal inhibitory drug concentration (IC50) was determined by using GraphPad Prism version 5.00 for Windows (GraphPad Software, CA).
Tam Y.T., Shin D.H., Chen K.E, & Kwon G.S. (2019). Poly(ethylene glycol)-block-poly(D,L-lactic acid) Micelles containing Oligo(lactic acid)8-Paclitaxel Prodrug: In Vivo Conversion and Antitumor Efficacy. Journal of controlled release : official journal of the Controlled Release Society, 298, 186-193.
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10

Resazurin-based Assay for Cell Viability

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Cell viability after HDAC inhibition was determined by a resazurin-based fluorimetric assay using the CellTiter-Blue® Cell Viability Assay (Promega, Madison, WI, USA) according to the manufacturer’s instructions. Non-fluorescent resazurin can be reduced to fluorescent resorufin by viable cells, and resorufin fluorescence can be determined (579Ex/584Em) [68 (link)]. Briefly, 100,000 cells were seeded in six-well plates and exposed to TSA or DMSO (control) for 6 h, 24 h, and 48 h. After that, 70 µL of CellTiter-Blue® was added to 375 µL medium per well and incubated at 37 °C for 1 h. Then, 100 µL was transferred into a 96-well plate and fluorescence was measured by a CytoFluor™ 2350 fluorometer (Millipore, Schwalbach, Germany). Cell viability was calculated relative to the corresponding DMSO controls.
Heppt M.V., Wessely A., Hornig E., Kammerbauer C., Graf S.A., Besch R., French L.E., Matthies A., Kuphal S., Kappelmann-Fenzl M., Bosserhoff A.K, & Berking C. (2022). HDAC2 Is Involved in the Regulation of BRN3A in Melanocytes and Melanoma. International Journal of Molecular Sciences, 23(2), 849.
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