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Prestoblue viability reagent

Manufactured by Thermo Fisher Scientific
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Sourced in United States, United Kingdom

PrestoBlue is a cell viability reagent that can be used to assess the metabolic activity of cells in culture. It is a resazurin-based solution that is reduced by metabolically active cells, resulting in a change in fluorescence and absorbance that can be measured.

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Lab products found in correlation

Erastin, by Bio-Techne (2 mentions) Victor 3v, by PerkinElmer (2 mentions) 96 well plate, by Corning (2 mentions) Penicillin, by Thermo Fisher Scientific (2 mentions) Streptomycin, by Thermo Fisher Scientific (2 mentions) Victor3v plate reader, by PerkinElmer (1 mentions) F500 plate reader, by Tecan (1 mentions) Complete medium, by Merck Group (1 mentions) Spectramax m2e, by Molecular Devices (1 mentions) Microplate reader, by Agilent Technologies (1 mentions) Apoptag fluorescein in situ apoptosis detection kit, by Merck Group (1 mentions) D300e digital dispenser, by Tecan (1 mentions) Victor x3, by PerkinElmer (1 mentions) Doxorubicin, by Merck Group (1 mentions) Amphotericin b, by Thermo Fisher Scientific (1 mentions) Paclitaxel, by Merck Group (1 mentions) Sodium azide, by Merck Group (1 mentions) 384 well plate, by Greiner (1 mentions) Envision multilabel reader, by PerkinElmer (1 mentions) Epoch microplate spectrophotometer, by Agilent Technologies (1 mentions)

Close spelling variants of this product

PrestoBlue (PB) Cell Viability Reagent PrestoBlue HS Cell Viability Reagent PrestoBlue Cell Viability Reagent PrestoBlue™ Viability Reagent reduction assay Prestoblue cell viability assay reagent PrestoBlue™ Cell Viability Reagent 1× PrestoBlue Cell Viability Reagent 10X solution PrestoBlue reagent PrestoBlue

31 protocols using prestoblue viability reagent

1

Immortalized Murine Kidney Cell Cultures

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Immortal murine mesangial cells and glomerular endothelial cells (Jieqing Biotech, Wuhan, China) were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum (8 (link), 33 (link)). Immortal murine proximal tubular epithelial cells (kindly provided by Dr. Cheng Pan, Wuhan University, China) were cultured in Dulbecco’s modified Eagle’s medium as described previously (34 (link)).
By using a HiTrap purification kit (GE Healthcare, Port Washington, NY, USA) (8 (link)), anti-dsDNA IgG was purified from the culture supernatants of murine WJ31 hydridoma clone (IgG2a; Jieqing Biotech). Control murine IgG (IgG2a; Jieqing Biotech) was verified by enzyme-linked immunosorbent assay (ELISA) to determine that there was no binding to DNA antigen (data not shown). Cells were starved routinely before the 2-day stimulation of anti-dsDNA IgG or control IgG (2 µg/ml). In some experiments, mesangial cells were stimulated with anti-dsDNA IgG (2 µg/ml) that was premixed with d-form (ALWPPNLHAWVP) or scrambled (PLPHNPWVLAAW) ALW peptide (1 µg/ml). The cell viability of three types of cells was measured using PrestoBlue viability reagent (Life Technologies, Carlsbad, CA, USA).
Wang P., Yang J., Tong F., Duan Z., Liu X., Xia L., Li K, & Xia Y. (2017). Anti-Double-Stranded DNA IgG Participates in Renal Fibrosis through Suppressing the Suppressor of Cytokine Signaling 1 Signals. Frontiers in Immunology, 8, 610.
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2

Chondrosarcoma cell lines response to mTOR inhibitors

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Chondrosarcoma cell lines JJ012, CH2879 and SW1353 counting seeded in previously optimized densities (JJ012 and SW1353 3000cells/well, CH2879 5000cells/well) were allowed to adhere overnight before treatment with increasing concentrations of mTOR inhibitors rapamycin and sapanisertib for 72 h. Cell viability was measured using presto blue viability reagent (Life-Technologies, Scotland, UK) and proliferation was measured using Hoechst staining as discussed previously. Experiments were performed in normoxic as well as hypoxic conditions by incubating cells in a MCO-19 M O2 /CO2 incubator (Panasonic) using nitrogen to simulate a 1% O2 environment and in combination with doxorubicin or cisplatin chemotherapeutic agents. Experiments were performed at least three times in triplicate.
Addie R.D., de Jong Y., Alberti G., Kruisselbrink A.B., Que I., Baelde H, & Bovée J.V. (2019). Exploration of the chondrosarcoma metabolome; the mTOR pathway as an important pro-survival pathway. Journal of Bone Oncology, 15, 100222.
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3

Rho-Inducible Cell Viability Assay

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Rho-expressing EcR 293 cell lines were seeded into a 96-well plate at 7.000 cells per well in 100 µl of DMEM AQmedia™ culture medium. Following cells adhesion, Rho induction was performed by replacing the culture medium by fresh medium containing Ponasterone A at 5 µg/ml. When needed, BCM was directly added to the wells at different concentrations (10, 20 or 50 µg/ml, final concentration) at the 12 h growth time point. Cells viability was assessed at each time point by using PrestoBlue™ Viability Reagent (life technologies ref: A13262) according to the manufacturer’s instructions. Briefly, 10 µl of the reagent were added directly to the culture medium in each well. Following 2 h incubation (37 °C, 5% CO2), the plate was shaken 5 min on a plate shaker then read with a Victor 3 V plate reader (PerkinElmer). Shortly after reading the plate, pictures were taken with a 16 million pixel camera by pipetting the colored media from the wells and pouring them into a new 96-well plate.
Moreau K., Surand J., Le Dantec A., Mosrin-Huaman C., Legrand A, & Rahmouni A.R. (2018). Recombinant yeast and human cells as screening tools to search for antibacterial agents targeting the transcription termination factor Rho. The Journal of Antibiotics, 71(4), 447-455.
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4

Cell Growth Assessment Across Breast Cancer Cell Lines

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The cell growths of MCF10A (2000 cells/well), MCF7 (1500 cells/well) and T47D (1500 cells/well), TamC3 (1500 cells/well) and TamR3 (2200 cells/well) were assessed in 96-well plates using Presto Blue Viability Reagent (Life Technologies Corporation, Carlsbad, CA, USA) according to the manufacturer’s protocol. Fluorescence was measured at excitation 535 nm and emission 612 nm using a Tecan F500 plate reader (Tecan, Grodig, Austria, GmbH). To detect cell growth for (1) siRNA silencing, we conducted reverse-transfection within a 96-well plate and analyzed data at different time points. (2) For showcasing the dependence of cells on SEMA3C, we used Plexin-B1 complex for growth, and exogenous SEMA3C (1 uM) was added to wells in presence of charcoal strip serum. (3) For IC50, cell growth inhibition was by B1SP and during synergy assay we allowed cells to attach to the wells of the 96-well plate overnight and then administered B1SP, Fulvestrant or tamoxifen the next day at the indicated concentrations in respective experiments (see results section).
Bhasin S., Dusek C., Peacock J.W., Cherkasov A., Wang Y., Gleave M, & Ong C.J. (2023). Dependency of Tamoxifen Sensitive and Resistant ER+ Breast Cancer Cells on Semaphorin 3C (SEMA3C) for Growth. Cells, 12(13), 1715.
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5

Proliferation of A375 Cell Lines

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To compare the proliferation of A375SPANX-KD and A375CTRL cell lines, 5 × 103 cells were seeded into 96-well plates in complete medium (DMEM supplemented with l-glutamine and 10% FBS) (Sigma-Aldrich) and incubated overnight. After 18 h, the cells were considered to be in T = 0. After further incubation for 24 h, cell viability was measured using PrestoBlue Viability Reagent (Life Technologies, Thermo Scientific, MA, USA) following the manufacturer’s instructions. Briefly, the medium was replaced with PrestoBlue diluted 1/10 in complete medium and the viability was measured after 2 h of incubation. The results shown are the means of three different experiments, and the error bars indicate the SEM.
Urizar-Arenaza I., Benedicto A., Perez-Valle A., Osinalde N., Akimov V., Muñoa-Hoyos I., Rodriguez J.A., Asumendi A., Boyano M.D., Blagoev B., Kratchmarova I, & Subiran N. (2021). The multifunctional role of SPANX-A/D protein subfamily in the promotion of pro-tumoural processes in human melanoma. Scientific Reports, 11, 3583.
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6

Evaluating Cell Viability by PrestoBlue

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Cells were plated in triplicate at 4 × 104 per well in 24-well culture plates, and protein expression was induced with doxycycline as before, for 48 hrs where appropriate. PrestoBlue Viability reagent (Invitrogen) was diluted 1:10 in normal growth medium and applied to cells for 20 mins at 37 °C, 5% CO2. Fluorescence was measured at 544 nm excitation and 590 nm emission with a Spectramax M2e (Molecular Devices) plate reader. Blank (no cell) well readings were averaged and subtracted from all experimental data points. Results were converted to a percentage viability relative to the respective clone uninduced reading, to allow comparison between clone types.
Chambers J.S., Brend T, & Rabbitts T.H. (2019). Cancer cell killing by target antigen engagement with engineered complementary intracellular antibody single domains fused to pro-caspase3. Scientific Reports, 9, 8553.
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7

Chondrosarcoma Cell Viability Assay

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Chondrosarcoma cells were counted using a Burker Turk counting chamber and seeded in optimized cell densities in 96 well plates. After attachment overnight cells were irradiated with increasing doses. Seventy-two hours after radiation cell viability was measured using PrestoBlue viability reagent (Invitrogen, Life-Technologies, Scotland, UK) according to the manufacturer’s instructions. Fluorescence was measured at 590 nM using a Wallac plate reader (Victor3V, 1420 multilabel counter, Perkin Elmer, the Netherlands). Experiments were performed three times in triplicate.
de Jong Y., Ingola M., Briaire-de Bruijn I.H., Kruisselbrink A.B., Venneker S., Palubeckaite I., Heijs B.P., Cleton-Jansen A.M., Haas R.L, & Bovée J.V. (2019). Radiotherapy resistance in chondrosarcoma cells; a possible correlation with alterations in cell cycle related genes. Clinical Sarcoma Research, 9, 9.
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8

Evaluating Cell Adhesion on Hyaluronic Acid

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6 well plates were coated with 50 μg/ml HA for 1 h, then washed 3 times with PBS and blocked in 2% BSA in PBS for 1 h. 50,000 cells per well were allowed to bind for 5 min and then washed 3 times with media. Bound cells were incubated with Presto-Blue viability reagent (Invitrogen) and quantified using a fluorescent plate reader (Molecular Devices).
Cieply B., Koontz C, & Frisch S.M. (2015). CD44S-hyaluronan interactions protect cells resulting from EMT against anoikis. Matrix biology : journal of the International Society for Matrix Biology, 48, 55-65.
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9

Evaluating Cell Adhesion on Hyaluronic Acid

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6 well plates were coated with 50 μg/ml HA for 1 h, then washed 3 times with PBS and blocked in 2% BSA in PBS for 1 h. 50,000 cells per well were allowed to bind for 5 min and then washed 3 times with media. Bound cells were incubated with Presto-Blue viability reagent (Invitrogen) and quantified using a fluorescent plate reader (Molecular Devices).
Cieply B., Koontz C, & Frisch S.M. (2015). CD44S-hyaluronan interactions protect cells resulting from EMT against anoikis. Matrix biology : journal of the International Society for Matrix Biology, 48, 55-65.
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10

Erastin-Induced Ferroptosis Modulation

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To induce ferroptosis, cells were incubated with erastin (Tocris, 5449) for 48 h, then cellular viability was evaluated with PrestoBlue viability reagent (Invitrogen, A13262) on a microplate reader (BMG labtech, Fluostar Optima). CDDO-Im (100 nM for LLC-PK1 and 10 nM for HK2 and MEF cells) or vehicle (dimethyl sulfoxide) were pretreated for 24 h to evaluate the contribution of NRF2 in ferroptosis. Cellular viability assays were repeated at least for 3 times, and representative experimental results are shown (all experiments showed the same trend). Relative viability was normalized to the respective erastin-free conditions. Sigmoidal nonlinear regression models were used to compute the regression fitting curves using GraphPad Prism.
Ide S., Ide K., Abe K., Kobayashi Y., Kitai H., McKey J., Strausser S.A., O’Brien L.L., Tata A., Tata P.R, & Souma T. (2022). Sex differences in resilience to ferroptosis underlie sexual dimorphism in kidney injury and repair. Cell reports, 41(6), 111610.
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