Celltiter fluor

Manufactured by Promega

Sourced in United States

CellTiter-Fluor is a fluorescent cell viability assay that measures the activity of a specific protease within live, metabolically active cells. It provides a quantitative measurement of cell viability in cell-based assays.

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Lab products found in correlation

Envision plate reader, by PerkinElmer (5 mentions) Celltiter glo, by Promega (4 mentions) Bright glo, by Promega (4 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (4 mentions) Synergy 4 microplate reader, by Agilent Technologies (3 mentions) One glo, by Promega (3 mentions) Spectramax plus 384, by Molecular Devices (2 mentions) Beta glo, by Promega (2 mentions) 384 well plate, by Corning (2 mentions) Protein g beads, by Avantor (2 mentions) Alamar blue, by Thermo Fisher Scientific (2 mentions) Indole 3 carbinol, by Merck Group (1 mentions) Ensight multimode plate reader, by PerkinElmer (1 mentions) Cytotox glo, by Promega (1 mentions) Cytotox one, by Promega (1 mentions) Wst 1, by Roche (1 mentions) Infinite 200 reader, by Tecan (1 mentions) One glo luciferase assay, by Promega (1 mentions) Hek293t 17, by American Type Culture Collection (1 mentions) Fetal bovine serum (fbs), by Cytiva (1 mentions)

Close spelling variants of this product

CellTiter-Fluor™ Cell Viability Assay kit CellTiter-Fluor cell viability CellTiter-Fluor reagent CellTiter-Fluor Viability Assay CellTiter-Fluor Assay CellTiter-Fluor Cell Viability Assay CellTiter-Fluor kit

41 protocols using celltiter fluor

Cell Viability and Caspase Assays

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Growth inhibition 50 (GI50) was determined by a fluorescence assay using 7-hydroxy-3H-phenoxazin-3-one 10-oxide (CellTiter-Blue, Promega) according to the manufacturer's protocol. Cell viability was determined by employing a fluorescence-based assay that relies in live-cell protease activity (CellTiter-Fluor, Promega) following the manufacturer's protocol. All fluorescence measurements were performed in a Synergy4 microplate reader (BioTek).
Caspase assays. Caspase-3 and -7 activity was assessed using the Apo-ONE caspase 3/7 assay (Promega) following the manufacturer's instructions with measurement of fluoresence emission in a Synergy4 microplate reader (BioTek). Caspase activity was normalized by the cell number determined by CellTiter-Fluor (Promega).

Pera B., Krumsiek J., Assouline S.E., Marullo R., Patel J., Phillip J.M., Román L., Mann K.K, & Cerchietti L. (2018). Metabolomic Profiling Reveals Cellular Reprogramming of B-Cell Lymphoma by a Lysine Deacetylase Inhibitor through the Choline Pathway. EBioMedicine, 28, 80-89.

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Cell Viability Assay of Indole-3-Carbinol

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Cells were plated in monolayer in black clear top 96-well plates with approximately 1.0 × 10⁴ cells/well in DMEM supplemented medium for 24 hours. Cells were then treated with indole-3-carbinol (Sigma-Aldrich, St. Louis, MO) (100 μM, 500 μM, 1 mM) or vehicle (DMSO) for 24 hours. The fluorescent cell viability assay CellTiter-FluorTM (Promega, Madison, WI) was used to assess cell fate following treatment. The fluorescence (excitation at 390 nm, emission at 460 nm) was measured using SpectraMax Plus 384 microplate reader (Molecular Devices).

Megna B.W., Carney P.R., Nukaya M., Geiger P, & Kennedy G.D. (2016). Indole-3-Carbinol Induces Tumor Cell Death: Function Follows Form. The Journal of surgical research, 204(1), 47-54.

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Luciferase Assay for PXR Activation by Indirubin

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Luciferase reporter assay to evaluate PXR activation by indirubin was performed using Human pregnane X receptor activation assay system following the manufacturer’s instructions (Puracyp Inc., Carlsbad, CA). Briefly, cells were seeded into 96-well plate and incubated overnight at 37 °C in 5% CO₂. Cells were then treated with DMSO (0.1%) or indirubin (100 nM). Rifampicin (100 nM) was used as positive control for PXR activation. Forty-eight hours post treatment, PXR activation was determined using ONE-Glo^TM Luciferase Assay System^TM and Celltiter-Fluor^TM (both from Promega, Madison, WI). Relative luminescence units (RLU) and relative fluorescence units (RFU) of each well were measured using EnSight Multimode Plate Reader (PerkinElmer Inc., Waltham, MA). PXR activation was then calculated by dividing normalized luciferase activity (RLU/RFU) of indirubin-treated condition by that of DMSO-treated control condition and was shown as fold activation relative to the vehicle control.

Tanaka Y., Uchi H., Ito T, & Furue M. (2019). Indirubin-pregnane X receptor-JNK axis accelerates skin wound healing. Scientific Reports, 9, 18174.

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Comparative Cytotoxicity and Proliferation Assays

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U373 cells and T98 cells were seeded at a density of 500 cells in 96-well plates and incubated for 24 hours. Each experiment was performed in triplicate. For the cell proliferation assays (WST-1 (Roche, Basel, Schweiz), CellTiter-GLO, CellTiter-Fluor (Promega), and Alamar Blue (Invitrogen)) as well as the cytotoxicity assays (CytoTox-GLO, CytoTox-One (Promega)) treatment effect was measured after 5 or 6 days of incubation. These time points were selected based on pilot time course experiments showing clear treatment effects at these points with control untreated cells still growing in the exponential growth phase. All assays were performed according to the manufacturers' instructions. Absorption, luciferase, or fluorescent signals were measured using the Infinite 200 reader (Tecan, Männedorf, Switzerland).
Results from the cytotoxicity assays are expressed as percentage of induced cytotoxicity compared to the nontreated control; error bars indicate standard deviation. Results from the viability assays are expressed as percentage of the nontreated control and error bars indicate standard deviation.

Kleijn A., Kloezeman J.J., Balvers R.K., van der Kaaij M., Dirven C.M., Leenstra S, & Lamfers M.L. (2016). A Systematic Comparison Identifies an ATP-Based Viability Assay as Most Suitable Read-Out for Drug Screening in Glioma Stem-Like Cells. Stem Cells International, 2016, 5623235.

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Cytotoxicity and mRNA Transfection Assay

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HeLa and HEK293T/17 cell lines (ATCC, Manassas, VA) were cultured in Dulbecco’s modified Eagle’s medium supplemented with 10% heat-inactivated FBS (Hyclone Laboratories Inc., Logan, UT) and 1% penicillin/streptomycin (Thermo Fisher, Federal Way, WA). All cultures were grown in 37°C incubators supplemented with 5% CO₂ and maintained following suppliers’ instructions. Cells were plated in white, clear-bottom 96-well plates at 4,000 cells per well and allowed to adhere overnight. Then, cells were administered LNPs encapsulating firefly luciferase mRNA. Cell viability results (CellTiter-Fluor^™, Promega), and luciferase expression data (ONE-Glo^™ Luciferase Assay, Promega) were collected 24 hours post-treatment with a microplate reader. Luminescent readout (in relative luminescence units, RLU) was normalized by cell counts commensurate with fluorescence (relative fluorescence units, RFU). RFU values were also compared between treated and untreated wells to determine cell viability.

Henderson M.I., Eygeris Y., Jozic A., Herrera M, & Sahay G. (2022). Leveraging Biological Buffers for Efficient Messenger RNA Delivery via Lipid Nanoparticles. Molecular pharmaceutics, 19(11), 4275-4285.

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Quantifying Host-Parasite Interactions

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Host cell and intracellular T. cruzi amastigote numbers were assessed as described with minor modifications [34 (link)]. Briefly, NHDF were plated at 1.5 x 10³ per well in 384 well plates and infected with MOI 1.25 for 2 hours before incubation in phenol-free media at the indicated glucose concentration. At 18 hpi, the indicated concentration of 2-DG was added, and at 66 hpi media was removed and host cell number and parasite number were assessed using 10 μL of CellTiter-Fluor (Promega) and 10 μL of Beta-Glo (Promega) per well, respectively.

Shah-Simpson S., Lentini G., Dumoulin P.C, & Burleigh B.A. (2017). Modulation of host central carbon metabolism and in situ glucose uptake by intracellular Trypanosoma cruzi amastigotes. PLoS Pathogens, 13(11), e1006747.

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Assessing Inhibition of Plasmodium Parasite

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Inhibition of P. berghei parasite load in hepatocytes was evaluated as previously described (Derbyshire et al., 2012 (link)). Briefly, 15,000 HepG2 or 8,000 HuH7 cells/well were seeded into 384-well plates in the presence or absence of compounds (0–100 μM) before infection with 3,000 P. berghei ANKA sporozoites. After 45 hrs, liver cell viability was assessed using CellTiter-Fluor (Promega) and parasite load was determined using Bright-Glo (Promega). The relative fluorescence and luminescence signal intensity of each well was normalized to the negative control (1% DMSO). Dose-response analysis was performed with GraphPad Prism.

Raphemot R., Eubanks A.L., Toro-Moreno M., Geiger R.A., Hughes P.F., Lu K.Y., Haystead T.A, & Derbyshire E.R. (2018). Plasmodium PK9 Inhibitors Promote Growth of Liver Stage Parasites. Cell chemical biology, 26(3), 411-419.e7.

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Cellular and In Vivo NAD Assays

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Example 3

Cellular assay: Test cells (for example, primary human fibroblasts) are seeded at 1×103 per well in 96-well tissue culture-treated dishes. Forty-eight hours later, cells are treated with either test compound, reference compound, or vehicle at a concentration of 1 mM. Cells are incubated at 37 C, 5% CO₂for twenty-four hours. Viability is assessed using CellTiter-Fluor™ (Promega, Madison, Wis.). Cells are then lysed and NAD(H) content is measured using the NAD/NADH-Glo™ assay (Promega, Madison, Wis.). NAD content is normalized to the number of viable cells and expressed relative to untreated control cells.

In vivo NAD and NAAD assay: Animals were dosed with 500 mg/kg of test compound or reference compound in vehicle. Results for Compound 2 are shown in FIGS. 1 and 2, which show that Compound 2 was orally available and boosted NAD and NAAD in murine liver and skeletal muscle, respectively, at the 8 hour time point.

US11279727B2. Anti-viral compounds and methods of use (2022-03-22). Metro International Biotech, LLC [US]. Inventors: Matthew Baevsky [US], Bruce Szczepankiewicz [US], Karen Lavery [US], David J. Livingston [US].

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Culturing JEG-3 Cells and HBMECs

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2D-cultured JEG-3 cells (2D SYNs; ATCC® HTB-36™, American Type Culture Collection, VA), human brain microvascular endothelial cells [HBMECs; (Coyne et al., 2007 (link))], LLC-MK2 cells (ATCC® CCL-7™; American Type Culture Collection, VA), and three-dimensional (3D) cultures of JEG-3 cells (3D SYNs) were grown as described previously (Silberstein et al., 2021 (link)). Experiments with 3D SYNs and 3D HBMECs were carried out between day 20–22 and day 4–6 after culture initiation, respectively. The number of viable cells was determined using the CellTiter-Fluor™ (for 2D SYNs) or the CellTiter-Glo™ 3D (for 3D SYNs) cell viability assays following manufacturer’s instructions (Promega, WI).

Silberstein E., Chung C.C, & Debrabant A. (2023). The transcriptome landscape of 3D-cultured placental trophoblasts reveals activation of TLR2 and TLR3/7 in response to low Trypanosoma cruzi parasite exposure. Frontiers in Microbiology, 14, 1256385.

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Assay for Trypanosoma cruzi Infection

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HFF were seeded at 1500 cells/well in 384 well black bottom plates (Corning). At 24h post plating, cells were infected with β-galactosidase expressing T. cruzi Tulahuen strain (moi 5) for 2hr, washed twice, and left in DMEM (2% FCS, 2 mM glutamine, 1 mM pyruvate). At 18hpi cells were treated with 0.3–10 μM of ELQ271. At 72 hpi HFF viability was measured in a fluorescence-based readout (CellTiter-Fluor, Promega) and T. cruzi was measured by luminescence-based readout (Beta-Glo reagent, Promega) using an Envision Plate Reader (PerkinElmer) as described [32 (link)]. The relative infection (RLU/RFU) was calculated and normalized to untreated control fitted by non-linear regression to high (= 100%) and low (= 0%) values using GraphPad Prism software.

Li Y., Shah-Simpson S., Okrah K., Belew A.T., Choi J., Caradonna K.L., Padmanabhan P., Ndegwa D.M., Temanni M.R., Corrada Bravo H., El-Sayed N.M, & Burleigh B.A. (2016). Transcriptome Remodeling in Trypanosoma cruzi and Human Cells during Intracellular Infection. PLoS Pathogens, 12(4), e1005511.

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