5x103 cells were seeded in 96-well plates overnight and treated with indicated compounds for 48 hours prior to addition of 40 μL 2 mg/mL MTT reagent (Promega) in PBS for 4 hours, and analyzed using SpectraMax M5 microplate reader (Molecular Devices, Sunnyvale, CA).
Mtt reagent
Manufactured by Promega
Sourced in United States
The MTT reagent is a colorimetric assay used to measure cell metabolic activity. It is a simple, quantitative, and reliable method for determining viable cell numbers.
Automatically generated - may contain errors
Lab products found in correlation
Thp 1, by American Type Culture Collection (3 mentions)
Hl 60, by American Type Culture Collection (3 mentions)
Celltiter 96 aqueous one solution cell proliferation assay, by Promega (3 mentions)
Microplate reader, by Bio-Rad (3 mentions)
Prism 6, by GraphPad (3 mentions)
Infinite m1000 pro plate reader, by Tecan (2 mentions)
Noble agar, by Merck Group (2 mentions)
Myd88, by Cell Signaling Technology (2 mentions)
Anti il 8 antibody, by Abcam (2 mentions)
α tubulin, by Cell Signaling Technology (2 mentions)
Myd88 knockdown constructs, by Merck Group (2 mentions)
Pd08959, by Selleck Chemicals (2 mentions)
Cd11b and cd14 antibodies, by Thermo Fisher Scientific (2 mentions)
Plvx ef1alpha ires mcherry vector, by Takara Bio (2 mentions)
Hct116, by American Type Culture Collection (2 mentions)
Sp600125, by Selleck Chemicals (2 mentions)
Sb203580, by Selleck Chemicals (2 mentions)
Oci aml3, by Leibniz Institute DSMZ (2 mentions)
Gapdh, by Santa Cruz Biotechnology (2 mentions)
Crystal violet, by Merck Group (2 mentions)
60 protocols using mtt reagent
1
Cell Viability Assay with MTT
2
Cytotoxic Effects of Amyloid-beta Species
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HT 22 cell was purchased from the Korean Cell line Bank (Seoul National University, Republic of South Korea) and cultured in the Dulbecco’s Modified Eagle’s medium (DMEM) (Thermo Fisher Scientific, USA) with 10% foetal bovine serum and 1% penicillin. The cytotoxic effect of synthesized Aβ1-42, Aβ4-42, and AβpE3-42 was investigated using the previously reported MTT assay protocol24 (link). Cultured HT22 cell (8 × 103 cells/well) was seeded on a 96-well plate. Each Aβ variant (10 μM) was prepared in starvation medium (0.5% foetal bovine serum and 1% penicillin in DMEM) and treated on the plate for 24 h at 37 C. 15 μL of MTT reagent (Promega, USA) was added to each well and incubated for additional 4 h at 37 C. Finally, 100 μl of solubilization solution (Promega, USA) was added for 30 min at 37 C. The absorbance was measured at 570 nm.
3
MTT Assay for Cell Viability
4
MTT Assay for Cell Viability
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After transfection, A549 cells were diluted to a certain concentration and paved at a 96-well plate at the density of 5 × 103 cells/well. Then the plate was incubated in a CO2 incubator at 37°C for 24, 48, and 72 h. Subsequently, each well was added with 20 μl of MTT reagent (Promega Corp., Madison, WI, U.S.A.) and cultured in an incubator for 4 h. Multifunctional microplate reader was employed to measure the optical density (OD) value of each well at 490 nm. The cell survival rate was calculated according to the OD value. Cell survival rate = (OD value of the experimental well − OD value of the blank well)/(OD value of the control well − OD value of the blank well) × 100%.
5
MTT Assay for Cell Viability and Proliferation
6
Assessing HSV1716 Cytotoxicity in HepG2-luc Cells
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HSV1716 toxicity in HepG2-luc cells was assessed using loss of light emission and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cell survival assays. HepG2-luc cells were plated out in the internal 6×10 grid of a 96-well tissue culture plate (Greiner Bio-One Ltd, Stonehouse, UK) at ∼5,000 cells per well. HSV1716 was added at increasing moi after 24 hours in culture in quadruplicate at least and, after a further 72 hours of incubation, the effect of the virus on light emissions and cell survival were determined. Light emission was detected after addition of 0.05 mL luciferase substrate to each well, and after 5 minutes of incubation at 20°C, light output was measured using a 1420 multilabel counter Victor 3 (Perkin-Elmer) in luminometer mode for 0.1 sec/well. Luciferin substrate was prepared by dissolving 1 g of D-luciferin, potassium salt (OZ Biosciences, Marseilles, France) in 66 mL of phosphate-buffered saline. Cell survival was assessed by addition of 0.01 mL of MTT reagent (Promega) to each well and the absorbance at 490 nm was determined after 1 hour of incubation at 37°C in 5% CO2. Killing curves for HSV1716 by loss of light emissions or MTT assay (GraphPad Prism version 4.02) were used to determine the ED50 (effective moi that kills 50% of the cells) for HSV1716.
7
Proliferation Assay of Endothelial Cells
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Cellular proliferation of isolated ECs was analyzed by MTT assay. 96-well plates were coated with fibronectin (BD), collagen (Corning, NY, USA), and gelatin (Sigma). Cells were plated at a cellular density of 1 × 104 cells per well in 100 μl of medium. After 24 h, 48 h, or 72 h incubation all wells received 20 μl of the MTT reagent (Promega, Madison, WI, USA) and were incubated for 2 h at 37 °C. Color intensity was measured in a plate reader (BMG Labtech) at 490 nm.
8
Cytotoxicity Assay Using MTT
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Approximately 104 cells were seeded into triplicate wells of 12-well plates and allowed to attach overnight. Either vehicle only or various concentrations of PD0325901, SB203580, LY294002, or paclitaxel (10 nM–10 μM, either alone or in combination) were then added to the cells. Growth was assessed by MTT [3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide] assay after 4 days of incubation. Briefly, after removal of medium, 1 ml of MTT reagent (0.5 mg/ml; Promega, Madison, WI, USA) was added to each well, and plates were incubated at 37°C for 30–60 min followed by the addition of 1 ml of acidic isopropanol and vigorous resuspension of the converted blue crystals. Absorbance of the suspension was measured at 595 nm with background subtraction at 650 nm. The effect of the inhibitor used was compared to vehicle-treated cells (taken as 100%).
9
MTT Assay for PC3 Cell Viability
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PC3 cells were seeded and transfected following the protocol described above in 96-well plates (4500 cells in 100 μl/well). After 24 h the supernatant was replaced by 100 μl of fresh growth medium (DMEM with 10% FCS). After further 24, 48 and 72 h of incubation 10 μl of MTT-reagent (Promega, Mannheim, Germany) were added to each well and incubated for 3 h at 37 °C. Absorption measurement at 570 nm was done with an Infinite M1000 Pro plate reader (Tecan, Männerdorf, Switzerland).
10
GSK3 Inhibitor Screening Library Protocol
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