Prestoblue

Manufactured by Thermo Fisher Scientific

Sourced in United States, United Kingdom, Germany, Japan, Canada, Australia, Switzerland, Ireland

PrestoBlue is a cell viability reagent used to assess the metabolic activity of cells. It is a resazurin-based solution that undergoes a color change when reduced by viable cells, allowing for the quantification of cell growth and proliferation.

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Lab products found in correlation

Prestoblue, by Agilent Technologies (10 mentions) Prestoblue, by Tecan (10 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (9 mentions) Synergy ht, by Agilent Technologies (9 mentions) Fluostar omega, by BMG Labtech (8 mentions) Fluostar omega fluorescence 556 microplate reader, by BMG Labtech (8 mentions) Infinite m200 pro, by Tecan (8 mentions) Infinite 200 pro, by Tecan (7 mentions) Celltiter glo, by Promega (6 mentions) Prism 7, by GraphPad (6 mentions) Microplate reader, by Agilent Technologies (6 mentions) Sh 9000lab, by Hitachi (6 mentions) Varioskan flash, by Thermo Fisher Scientific (5 mentions) Propidium iodide, by Thermo Fisher Scientific (5 mentions) Fetal bovine serum (fbs), by Merck Group (5 mentions) Infinite m200, by Tecan (5 mentions) Presto blue, by PerkinElmer (5 mentions) Calcein am, by Thermo Fisher Scientific (5 mentions) 96 well plate, by Corning (5 mentions) Dimethyl sulfoxide (dmso), by Merck Group (5 mentions)

Spelling variants of this product

PrestoBlue Cell Viability Reagent (642 citations) PrestoBlue solution (9 citations) PrestoBlue HS (9 citations) PrestoBlue™ Viability Reagent reduction assay (3 citations) PrestoBlue Cell Viability Assay kit (3 citations) PrestoBlue Cell Viability Reagent 10X solution (2 citations) PrestoBlue resazurin dye (2 citations) PrestoBlue (PB) Cell Viability Reagent (2 citations) PrestoBlue™ Cell Viability Reagent 1× (2 citations) PrestoBlue HS cell viability dye (1 citations)

489 protocols using prestoblue

Cell Viability Assessment via Presto Blue

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The 1 × 10⁴ cells were seeded in 96-well plates and incubated at 37 °C, 5% CO₂ for 24 h. Cell viability was assessed using Presto Blue reagent: cells were washed with PBS and incubated during 30–45 min with Presto Blue solution (1:10 prepared with Presto Blue, Thermo Fisher, Whatham, MA, USA, and DMEM medium). After the incubation time, fluorescence was measured in Synergy Mx, Biotek Instruments Inc (Winooski, VT, USA) (emission: 560 nm; excitation: 590 nm).

Dionísio M.R., Vieira A.F., Carvalho R., Conde I., Oliveira M., Gomes M., Pinto M.T., Pereira P., Pimentel J., Souza C., Marques M.M., Duval da Silva V., Barroso A., Preto D., Cameselle-Teijeiro J.F., Schmitt F., Ribeiro A.S, & Paredes J. (2020). BR-BCSC Signature: The Cancer Stem Cell Profile Enriched in Brain Metastases that Predicts a Worse Prognosis in Lymph Node-Positive Breast Cancer. Cells, 9(11), 2442.

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Breast Cancer Cell Viability Assay

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Cell viability in varying concentrations of the Au@peptide NPs was assessed using PrestoBlue cell viability assay. Human breast adenocarcinoma (MDA-MB-231) and lung fibroblast (IMR-90) cells were seeded in a 96-well flat bottom microplate (BioLite Microwell Plate, Fisher Scientific, Waltham, MA). For IMR-90, 6.0 × 10³ cells per well were plated, and for MDA-MB-231, 5.6 × 10³ cells per well were plated in 100 μL of DMEM culture media. The cells were allowed to grow for 24 h at 37 °C and 5% CO₂ in a humidified incubator. At confluency, cells were dosed with 0, 1, 2, 5, 10, or 50 nM of Au@peptide NPs (in triplicate). Following the administration of the NPs, cells were incubated for 48 h at 37 °C under 5% CO₂. After each period of incubation, PrestoBlue (Life Technologies, Carlsbad, CA) was used as an indicator of cellular toxicity; 11 μL of PrestoBlue was added to each well and incubated for 1 h at 37 °C under 5% CO₂. The 96-well plate was then analyzed using a multimode plate-reader BioTek Microplate Reader (BioTek U.S., Winooski, VT) at 560 / 590 nm wavelength. The percentage of surviving cells was calculated as a normalized ratio of the fluorescence intensity between cells treated with AuNPs and media alone.

Huang R.H., Nayeem N., He Y., Morales J., Graham D., Klajn R., Contel M., O’Brien S, & Ulijn R.V. (2021). Self-Complementary Zwitterionic Peptides Direct Nanoparticle Assembly and Enable Enzymatic Selection of Endocytic Pathways. Advanced materials (Deerfield Beach, Fla.), 34(1), e2104962.

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Evaluating Cellular Toxicity of Peptides

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Human fetal lung fibroblast (IMR90) and human clear cell renal cell carcinoma cells (Caki-1) were seeded in 96-well flat bottom microplate (BioLite Microwell Plate, Fisher Scientific, Waltham, MA). For IMR90 we used 6.0 X 10³ cell per well and for Caki-1 we used 5.6 X 10³ cells per well in 90μL of complete phenol red free cell culture media. The cells were allowed to grow for 24hours at 37°C and 5% CO₂ in a humidified incubator. 10mM peptide solutions were prepared in phosphate buffer saline, pH adjusted to 7.4 using NaOH or HCl and sonicated for 10minutes. Then 10μL of each sample was added into wells containing 90μL of media (in triplicate). Following the administration of the peptides, cells were incubated for 72 hours at 37°C under 5% CO₂. After each period of incubation, Presto Blue (Life Technologies, Carlsbad, CA) was used as an indicator of cellular toxicity; 11μL of Presto Blue was added to each well and incubated for 1 hours at 37°C under 5% CO₂. The 96-wells plate was then analyzed using a multi-mode plate-reader BioTek Microplate Reader (BioTek U.S., Winooski, VT) at 530nm and 590nm wavelength. The percentage of surviving cells was calculated as a normalized ratio of the fluorescence intensity between cells treated with media and phosphate buffer saline alone.

Son J., Kalafatovic D., Kumar M., Yoo B., Cornejo M.A., Contel M, & Ulijn R.V. (2019). Customizing Morphology, Size, and Response Kinetics of Matrix Metalloproteinase-responsive Nanostructures by Systematic Peptide Design. ACS nano, 13(2), 1555-1562.

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Cell Viability Assay with PrestoBlue™

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Cell viability experiments were performed using PrestoBlue™ (Invitrogen, U.S.A.) cell viability reagent. PrestoBlue™ cell viability reagent is
a ready-to-use reagent for rapid evaluation of the viability and proliferation of a wide range of cell types. Measurement of the bystander cell viability
was conducted by seeding the cells at a density of 3 × 10³ cells/well in 96-well plates. This density was found to be optimal to achieve the desired confluence
at the end of the exposure period for both MCF-7 and hFOB 1.19 cell lines. After 24 and 48 hours of incubation with ICCM, the samples were washed twice with PBS
and then PrestoBlue™ reagent was added in each well. After 2 hours incubation at 37°C and 5% CO₂, the absorbance from colorimetric changes of PrestoBlue™ reaction with viable
cells were measured using microplate reader (Varioskan Flash, Thermo Scientific) at an excitation wavelength of 560 nm and an emission wavelength of 590 nm.

Zainudin NH. M., R. A, & W. N. R. (2020). Bystander Effect Induced in Breast Cancer (MCF-7) and Human Osteoblast Cell Lines (hFOB 1.19) with HDR-Brachytherapy. Journal of Biomedical Physics & Engineering, 10(3), 319-328.

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DHT Stimulation and Cell Growth Assay

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For the DHT stimulation experiment, 1 × 10e4 cells, and for the non-DHT experiment, 2 × 10e3 cells were seeded into 48-well plates in K-SFM. The next day, alamarBlue or prestoBlue (Invitrogen/Thermo Fisher Scientific) premixed with hormone-stripped medium supplemented with 0, 1, 10, or 100 nM DHT or with K-SFM was changed to the cells for 2 h (alamarBlue used for DHT stimulations) or 30 min (prestoBlue used for K-SFM) on each day of measurement (different wells each day) for 7 days. Cell growth was measured in six (K-SFM growth curves) or eight (DHT-stimulations) replicates. Fluorescence was measured with an EnVision 2104 multilabel reader (PerkinElmer) with excitation 570 nm and emission 585 nm. Growth curve data were normalized to day 1 results, and differences in growth between conditions were assessed with one-way ANOVA and Tukey’s post hoc test with GraphPad Prism v5.02.

Kukkonen K., Autio-Kimura B., Rauhala H., Kesseli J., Nykter M., Latonen L, & Visakorpi T. (2022). Nonmalignant AR-positive prostate epithelial cells and cancer cells respond differently to androgen. Endocrine-Related Cancer, 29(12), 717-733.

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Determining Non-Toxic Cell Viability

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The optimal non-toxic concentrations of the AM extract and of the AC used in this study were determined by using the PrestoBlue® reagent (Invitrogen, USA). DPCs at a density of 5 × 10³ cells/well in 96-well plates, were cultured overnight in complete DMEM. The cells were separately treated with various concentrations of the AM extract (0–40 μg/mL) and of the AC (0–40 μM). Cell viability was measured after an incubation with PrestoBlue® in Roswell Park Memorial Institute (RPMI) medium (Invitrogen, USA) at 37°C, for 48 h. If there were viable cells, PrestoBlue® would change from a blue to a purple-pink color, that could be spectrophotometrically detected at 570 nm.

Prugsakij W., Numsawat S., Netchareonsirisuk P., Tengamnuay P, & De-Eknamkul W. (2023). Mechanistic synergy of hair growth promotion by the Avicennia marina extract and its active constituent (avicequinone C) in dermal papilla cells isolated from androgenic alopecia patients. PLOS ONE, 18(4), e0284853.

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Cell Viability Evaluation via PrestoBlue Assay

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Cell viability analysis was carried out using PrestoBlue (Invitrogen, MA, USA) cell viability reagent according to the manufacturer’s instructions. In brief, culture medium was removed and cells washed with phosphate buffered saline (PBS, Merck, MA, USA) containing calcium and magnesium chloride (referred to hereon as PBS^+/+). Following washing, one well of the dark control cells was exposed to cell lysis buffer (RIPA Lysis and Extraction Buffer, Thermo Scientific, MA, USA) for 5 min at room temperature to act as a positive control. Next, 120 µL of 1:10 dilution of PrestoBlue (Invitrogen, MA, USA) cell viability reagent and PBS^+/+ was added to each exposed well. Cells were then incubated at 37 °C and 5% CO₂ for 20 min to allow the assay to develop. Following incubation, 100 µL of the developed reagent was transferred to a solid white 96-well assay plate and fluorescence was read using a multi-modal plate reader (Ex 520 nm/Em 580 nm; GloMax Explorer, Promega, WI, USA).

Anderson G., McLeod A., Bagnaninchi P, & Dhillon B. (2022). Quantitative action spectroscopy reveals ARPE19 sensitivity to ultraviolet radiation at 350 nm and 380 nm. Scientific Reports, 12, 14223.

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GET-FYLIR Cytotoxicity Assay in Cells

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Cells were plated in 96-well plates (2.5 × 10⁴ cells/well) and treated with GET-FYLIR (0–50 μm) for 24 h. Cells were also treated with GET-Scrambled as a control. After treatment, cells were washed with PBS and then incubated for 15 min at 37 °C with a 100-μl Presto Blue solution (10% Presto Blue, Invitrogen in Hanks' balanced salt solution, Sigma). Change in the fluorescence was measured using a plate reader with the excitation/emission wavelengths set at 560/590 nm (Infinite® 200 PRO, TECAN), with untreated cells used as a control. All data sets were combined for the statistical analysis. A Sidak's multiple comparisons test was applied. N refers to the number of biological replicates.

Spiliotopoulos A., Blokpoel Ferreras L., Densham R.M., Caulton S.G., Maddison B.C., Morris J.R., Dixon J.E., Gough K.C, & Dreveny I. (2018). Discovery of peptide ligands targeting a specific ubiquitin-like domain–binding site in the deubiquitinase USP11. The Journal of Biological Chemistry, 294(2), 424-436.

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Optimizing hiPSC Culture Conditions

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The hiPSC line 19c3 (p20–p60), derived from a healthy male, was used for medium variable optimization. Prior to assay, cells were grown to 75% confluence after 4 days of culture as above. Cells were dissociated with TrypLE (Gibco, 12604013) for 3 min at 37°C and resuspended in DMEM/F12, transferred to a 15-mL conical tube (Falcon), and centrifuged at 200 × g for 3 min (Sorvall ST40). The pellet was resuspended in DMEM/F12 and diluted to 1 × 10⁵ cells mL⁻¹, and 10,000 cells were plated per well in Matrigel (1:800)-coated 12-well plates (Greiner) in the medium to be tested along with 2 μM thiazovivin for the first 24 h. Media were changed daily and cells were grown for 6 days. This lower than normal seeding density was used to allow the discovery of factors detectable only under more extreme conditions and therefore provide data on the robustness of the formulation. Cell growth was then assessed using PrestoBlue (Invitrogen, A13262), by adding 100 μL of PrestoBlue to the 1 mL of existing medium in each well and incubating for 2 h at 37°C. Fluorescence (560 nm excitation, 590 nm emission) was then measured using a Varioskan LUX (Thermo Scientific) plate reader with “top read” function.

Kuo H.H., Gao X., DeKeyser J.M., Fetterman K.A., Pinheiro E.A., Weddle C.J., Fonoudi H., Orman M.V., Romero-Tejeda M., Jouni M., Blancard M., Magdy T., Epting C.L., George AL J.r, & Burridge P.W. (2020). Negligible-Cost and Weekend-Free Chemically Defined Human iPSC Culture. Stem Cell Reports, 14(2), 256-270.

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Viability Assessment of Dental Pulp Stem Cells

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For live/dead imaging, DPSCs were grown on sterile cover slides 24 × 24 mm and cultured for 5 days in FBS or PL media. After formalin fixation, the cells were stained with a solution of Calcein AM-EthD-III (Biotium, Hayward) according to the kit indications. Epifluorescent signal was detected and relative images were acquired through SP5 microscope (Leica, Wetzlar). Metabolic activity of DPSCs was assessed by PrestoBlue (Molecular Probes, Eugene) assay. Briefly, cryopreserved DPSCs were thawed and resuspended in α-MEM containing FBS at 10% or PL at 5%, 2%, and 1%. Cells were seeded at 2500 cell/well in one 96-well plate for each analysed time point; more than 6 wells were replicated for each condition. An aliquot of fresh medium was provided to the plates that were cultured for more than 4 days. The PrestoBlue reagent was incubated as supplier instruction to a final volume of 100 μL; after 2 hours the collected supernatant was measured for absorbance with Multiskan GO (Thermo Fisher, Waltham) spectrophotometer (570–600 nm). An additional control was set using human serum (HS) 10%. For viable cell counts, the samples were trypsinized after 3 days or 3 weeks and measured by ADAM-MC (AlphaMetrix, Rödermark) system to evaluate cell membrane permeability by dye exclusion.

Marrazzo P., Paduano F., Palmieri F., Marrelli M, & Tatullo M. (2016). Highly Efficient In Vitro Reparative Behaviour of Dental Pulp Stem Cells Cultured with Standardised Platelet Lysate Supplementation. Stem Cells International, 2016, 7230987.

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