Proliferation was measured in quadruplicate by MTT Cell Proliferation Assay (ATCC) following the manufacturer’s protocol. Apoptosis was measured by flow cytometry after staining with Annexin V-APC (Life Technologies) and PI (Sigma) in 10 mM HEPES at pH 7.4, 140 mM NaCl, 2.5 mM CaCl2. For cell cycle analysis, cells were washed once in PBS, fixed in 70% ethanol at 4°C, washed again in PBS and resuspended in 0.1% (v/v) Triton X-100 in PBS, containing 200 μg/ml DNase-free RNase A and 20 μg/ml PI. Samples were incubated at 37°C for 15 min before analysis on a Becton-Dickinson FACScan using FlowJo (Tree Star, Inc).
Mtt cell proliferation assay
Manufactured by American Type Culture Collection
Sourced in United States
The MTT Cell Proliferation Assay is a colorimetric assay used to measure the metabolic activity of cells. It is a quantitative method for determining the number of viable cells in proliferation or cytotoxicity experiments.
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Lab products found in correlation
Plasmocin prophylactic, by InvivoGen (2 mentions)
Trypan blue solution, by Merck Group (2 mentions)
Fetal bovine serum (fbs), by Atlanta Biologicals (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Annexin 5 apc, by Thermo Fisher Scientific (1 mentions)
Facscan, by Tree Star (1 mentions)
Spectramax m5, by Molecular Devices (1 mentions)
96 well microplate, by Corning (1 mentions)
96 well flat bottomed microtiter plates, by BD (1 mentions)
Amphotericin, by Thermo Fisher Scientific (1 mentions)
Estradiol, by R&D Systems (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Caco 2 cell, by American Type Culture Collection (1 mentions)
Deadend fluorometric tunel assay kit, by Promega (1 mentions)
Afuresertib, by Cayman Chemical (1 mentions)
Gw3965, by R&D Systems (1 mentions)
Dnase, by Thermo Fisher Scientific (1 mentions)
Sw620 cells, by American Type Culture Collection (1 mentions)
Immun blot pvdf membrane for protein blotting, by Bio-Rad (1 mentions)
28 protocols using mtt cell proliferation assay
1
Assessing Cell Proliferation and Apoptosis
2
MTT Assay for T Cell Proliferation
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Proliferation of cultured primary CD4 T cells was measured using the MTT Cell Proliferation Assay (ATCC) following manufacturer’s instructions. Briefly, 80,000 cells were plated in 90 μl of media in flat bottom plates in the presence or absence of T-activator beads. After 24h, 10 μl of MTT reagent was added to each well. Plates were incubated at 37°C in a 5% CO2 incubator for 4 h. 100 μl of detergent reagent was added to each well and plates were incubated in the dark at room temperature overnight. Absorbance was measured at 570 and 650 nm. During analysis, the 650 nm absorbance and media alone controls were subtracted from each sample.
3
MTT Cell Viability Assay Protocol
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3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay performed by MTT Cell Proliferation Assay (cat # 30–1010K) following the manufacturer’s protocol (ATCC, Manassas, VA, USA) was used to determine the cellular viability [27 (link)]. Finally, the number of viable cells in each well was determined by a microplate reader (SpectraMax M5, Molecular Devices, Sunnyvale, CA, USA) at 570 nm. Cell viability was presented as percent viability relative to vehicle-treated control.
4
Cell Viability Assessment by MTT Assay
5
MTT Cell Proliferation Assay
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Cell proliferation was determined using the MTT Cell Proliferation Assay (ATCC), according to manufacturer’s directions.
6
Evaluating Botanical Extracts' Impact on Hep G2 Cell Viability
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Percentage cell viability was determined using MTT Cell Proliferation Assay according to the manufacturer’s protocol (ATCC, Manassas, VA). Hep G2 cells were grown in 96-well plates at a density of ~10,000 cells/well. The cells were exposed for 72 hr to 5 mM fructose with or without the botanical extracts from 0 to 500 μg/mL. Extracts from Lot #1 of Angelica archangelica, Scutellaria baicalensis, Petroselinum crispum, and Garcinia mangostana were used. Experiments were done in triplicates and the absorbance was read at 570 nm.
7
Cytotoxicity Evaluation of SPACE Peptide
8
HPASMC Proliferation Assay
9
Cell Proliferation Quantification
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MTT cell proliferation assay (ATCC, Manassas, VA) kit was used to quantify cell proliferation, following manufacturer’s protocol.
10
MTT Assay for Cytotoxicity Evaluation
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The MTT cell proliferation assay (catalog no. 30-1010K-ATCC; Manassas, VA, USA) was used to measure the cell proliferation rate. The assay was performed to study the cytotoxicity effect of ISRAA on the cell lines according to the manufacturer’s instructions. Cell suspension was harvested by centrifugation and resuspended at 1×106/ml. Serial dilutions of ISRAA in culture medium were prepared from 50 μg to 10 pg/ml. The cell suspension (100 μl) was plated in each well in triplicate and three control wells of medium alone (to provide the blanks for absorbance readings) were included, in addition to another three wells with cell suspension alone as a negative control.
Each serial dilution of ISRAA protein (10 μl) was added to each well in triplicate. Plates were incubated at 37°C in 5% CO2 for 24, 48 and 72 h. Later on, 10 μl of 5 mg/ml MTT reagent was added to each well, including the controls and incubated for 4 h at 37°C. Following this, 100 μl of the MTT solvent (acidic isopropanol 0.04 M HCl in absolute isopropanol) was added to all wells including controls. Plates were covered with tinfoil and agitated on an orbital shaker for 15 min. Finally, the absorbance at 690 nm was recorded and the cytocide rate was calculated from the below formula: Cytocide rate (%) = (OD control − OD experiment)/OD control × 100.
Each serial dilution of ISRAA protein (10 μl) was added to each well in triplicate. Plates were incubated at 37°C in 5% CO2 for 24, 48 and 72 h. Later on, 10 μl of 5 mg/ml MTT reagent was added to each well, including the controls and incubated for 4 h at 37°C. Following this, 100 μl of the MTT solvent (acidic isopropanol 0.04 M HCl in absolute isopropanol) was added to all wells including controls. Plates were covered with tinfoil and agitated on an orbital shaker for 15 min. Finally, the absorbance at 690 nm was recorded and the cytocide rate was calculated from the below formula: Cytocide rate (%) = (OD control − OD experiment)/OD control × 100.
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