Heat inactivated FBS (fetal bovine serum), antibiotic–antimycotic, Dulbecco’s phosphate buffered saline (DPBS), high glucose (hg) DMEM, M199, Endothelial Cell Growth Medium (ECM) and Geltrex™ LDEV-Free Reduced Growth Factor Basement Membrane Matrix and NuPage 4–12% bis-Tris Gel, were purchased from Gibco-Life Technologies (Carlsbad CA, USA). Trypsin–EDTA solution1X, Dimethyl sulphoxide (DMSO), lipopolysaccharide (LPS) (E. coli 055:B5), Protein Assay Kit TP0300, In Vitro Toxicology Assay Kit and Cell Growth Determination Kit MTT based were purchased from Sigma-Aldrich (St. Louis, MO, USA). Pericyte Growth Medium was purchased from Promocell (Heidelberg, Germany). NucleoSpin RNA kit was purchased from Macherey-Nagel GmbH & Co. KG (Düren, Germany) RT2 strand kit, RT2 Sybr green fluor qPCR master mix were from Qiagen (Hilden, Germany). Porcine Cytokine/Chemokine Magnetic Bead Panel kit, Milliplex Map Kit EMD was purchased by Millipore Corporation (Billerica, MA, USA). Super Signal West Pico Chemiluminescent Substrate was from Pierce Biotechnology, Inc. (Rockford, IL, USA).
Cell growth determination kit mtt based
Manufactured by Merck Group
Sourced in United States
The Cell Growth Determination Kit MTT based is a laboratory product designed to measure cell viability and proliferation. The kit utilizes the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, a colorimetric method for assessing cell metabolic activity. The core function of the kit is to provide a quantitative assessment of cell growth and cytotoxicity in cell culture experiments.
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Lab products found in correlation
Nucleospin rna kit, by Macherey-Nagel (1 mentions)
Trypsin edta solution 1x, by Merck Group (1 mentions)
Geltrex ldev free reduced growth factor basement membrane matrix, by Thermo Fisher Scientific (1 mentions)
Rt2 sybr green fluor qpcr mastermix, by Qiagen (1 mentions)
Porcine cytokine chemokine magnetic bead panel kit, by Merck Group (1 mentions)
Pericyte growth medium, by PromoCell (1 mentions)
Dimethyl sulphoxide, by Merck Group (1 mentions)
In vitro toxicology assay kit, by Merck Group (1 mentions)
Antibiotic antimycotic, by Thermo Fisher Scientific (1 mentions)
Trypsin edta 0.25, by Merck Group (1 mentions)
Spectrophotometric microplate reader, by Beyotime (1 mentions)
Annexin 5 fitc pi apoptosis detection kit, by BD (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Dactolisib, by Santa Cruz Biotechnology (1 mentions)
Su1498, by Santa Cruz Biotechnology (1 mentions)
Dmem nutrient mixture f 12 ham, by Merck Group (1 mentions)
Microplate reader, by Beckman Coulter (1 mentions)
Dimethyl sulfoxide (dmso), by Thermo Fisher Scientific (1 mentions)
Curcumin, by Merck Group (1 mentions)
Thermo fisher multiskan fc, by Thermo Fisher Scientific (1 mentions)
17 protocols using cell growth determination kit mtt based
1
Pericyte Cell Culture and Characterization
2
Cytotoxicity and Apoptosis Evaluation
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The cytotoxicity of the trans-fected cells was measured using the Cell Growth Determination kit MTT based (Sigma-Aldrich; Merck KGaA). The cells were cultured and seeded into 96-well plates at a density of 2×103 cells per well. Trypsin/EDTA 0.25% (Sigma-Aldrich; Merck KGaA) was then added to each well to generate a single cell suspension. Following incubation for 4 h at 37°C with 5% of CO2, 150 µl MTT solvent (4 mM HCl, 0.1% NP40 in isopropanol) was added to each well and incubated at 7°C for 3 h for conversion into formazan. Finally, the optical density (OD) was determined at a wavelength of 450 nm with a spectrophotometric microplate reader (Beyotime Institute of Biotechnology).
Cell apoptosis was tested using the Annexin V-FITC/PI Apoptosis Detection kit (BD Biosciences) according to the manufacturer's instructions. The transfected cells were collected and Annexin V-FITC and propidium iodide (PI) were added to incubation at 37°C in the dark for 15 min. Cell apoptosis was performed and displayed using flow cytometry.
Cell apoptosis was tested using the Annexin V-FITC/PI Apoptosis Detection kit (BD Biosciences) according to the manufacturer's instructions. The transfected cells were collected and Annexin V-FITC and propidium iodide (PI) were added to incubation at 37°C in the dark for 15 min. Cell apoptosis was performed and displayed using flow cytometry.
3
MTT Assay for Cell Viability
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MTT assay was performed by Cell Growth Determination Kit MTT Based (Sigma-Aldrich). MTT solution was added in an amount equal to 10% of the culture volume and incubated for 3 to 4 h. The resulting MTT formazan crystals read as absorbance at a wavelength of 570 nm were spectrophotometrically measured.
4
Cell Proliferation Assay with Inhibitors
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Dulbeccos Modified Eagle Medium (DMEM)/Nutrient Mixture F-12 Ham, Dimethyl Sulfoxide (DMSO), Ethylenediaminetetraacetic Acid (EDTA), Cell Growth Determination Kit MTT based [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] were purchased from Sigma-Aldrich (Germany), Fetal Bovine Serum (FBS), Penicillin/Streptomycin antibiotics, Trypsin, Phosphate-buffered saline (PBS) were obtained from Gibco by Life TechnologiesTM, Cell Lysis Buffer was provided by Invitrogen, Life Tehnologies, USA.
SU1498 (VEGFR2 selective inhibitor) and Dactolisib (dual PI3K/mTOR inhibitor) were acquired from Santa Cruz Biotechnologies, AG1433 (PDGFR-β and a weak inhibitor of angiogenesis and VEGFR2) were obtained from Sigma-Aldrich.
SU1498 (VEGFR2 selective inhibitor) and Dactolisib (dual PI3K/mTOR inhibitor) were acquired from Santa Cruz Biotechnologies, AG1433 (PDGFR-β and a weak inhibitor of angiogenesis and VEGFR2) were obtained from Sigma-Aldrich.
5
Evaluating the Interplay of CD40 and Cyclosporine A on RCC Proliferation
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The effect of CD40-ligation and cyclosporine A on RCC cell proliferation was analyzed with MTT assay, using the cell growth determination kit MTT based (Sigma). Briefly, cultured RCCs were digested with 0.05% trypsin and then were cultured in serum-free medium in 96-well culture plates (100 μL per well). RCCs were pre-incubated with cyclosporine A (5 μM) for 18 h and/or stimulated with sCD40L with the enhancer or the enhancer alone for 24 or 48 h and then supplemented with 10 μL MTT (5 mg/mL) and incubated for another 4 h. The supernatant was then discarded by aspiration and the RCC preparation was shaken with 100 μL MTT solvent for 10 min, before the OD value was measured at 570 nm.
6
MTT Assay for Cell Proliferation
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3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay was performed for cell proliferation analysis using cell growth determination kit (MTT based) (Sigma-Aldrich). A549 and H1299 cells with or without AC074117.1 silencing were treated with MTT (10 μL) for 5 h. Then, the cells were incubated with DMSO (150 μL) for 10 min. Absorbance at 450 nm was detected by applying a microplate reader (Beckman Coulter, CA, USA).
7
Cytotoxicity of OXA and Curcumin in HCT116 Cells
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To explore the effect of OXA on the growth of HCT116 and HCT116/OXA cells, HCT116 and HCT116/OXA cells were cultured in RPMI1640 containing 0.5, 1, 2, 4, 8, 16 or 32 μM OXA for 48 hrs. To explore the effect of curcumin on HCT116/OXA cell growth, the cells were cultured with 1, 2, 4, 8, 16, 32 or 64 μM curcumin (MedChemExpress, Monmouth Junction, NJ, USA) for 48 hrs. The cytotoxicity of OXA, curcumin and combination of treatment were assessed by using cell growth determination kit (MTT based) (Sigma-Aldrich, St. Louis, MO, USA) according to the manufacturer’s instruction. In brief, HCT116 and HCT116/OXA cells (in the logarithmic growth phase) were seeded into 96-well plate with 3.0×103 cells per well. MTT reagent was added into each well and incubated for 4 hrs. Then, 150 μl dimethyl sulfoxide (DMSO, Invitrogen, Carlsbad, CA, USA) was added into each well for 10 mins. The absorbance at 570 nm (A570) was measured by Thermo Fisher Multiskan FC (Thermo Fischer Scientific, Waltham, MA, USA).
8
Cell Viability Assay of Transfected Cells
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Cell viability was determined using the "Cell growth determination kit MTT based" (Sigma-Aldrich, Saint Quentin Fallavier, France). The MTT assay was performed in quadruplicate in 48-well plates. To this end, 55,000 MRC5-V2 cells were seeded in 48-well plates in DMEM supplemented with 10% of serum. The next day, the cells were transfected. 24 h after transfection, the cells were incubated for 3h at 37°C in the presence of 500 µL of medium containing MTT reagent (0.5 mg/mL). The enzymatic reaction was stopped by removing the culture medium and by adding DMSO. The absorbance was then measured at 570 nm (and 690 nm) and the percentage of cell viability was calculated relative to untreated cells (the control "untreated cells" gives the value of 100% viability). The MTT assay was performed in triplicates; the last well of the quadruplicate was used to determine the transfection efficiency since it is not possible to perform the MTT and luciferase assays on the same sample.
9
Microglia Cell Viability Assay
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Cell viability was assessed by MTT reagent. MTT assays were performed according to the manufacturer's protocol from the Cell Growth Determination Kit MTT Based (Sigma, St. Louis, MO, USA). BV2 microglia were seeded in triplicate at a density of 2 × 10 4 cells/well on a 96-well plate. After the supernatant were removed, the cells were incubated with MTT solution (0.5 mg/ mL) at 37 °C for 4 h in an incubator. Then, the MTT solution was discarded, and an equal volume of DMSO was added to dissolve the formazan. The absorbance was measured at a wave length of 570 nm.
10
Dose-Dependent Cell Survival Assay
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Cells were seeded at subconfluent levels (<50% confluence) in 96-well tissue culture plates and treated with increasing concentrations of MS-444 [21 (link)] for 48 hours at 37°C. Cell survival was assayed using the MTT-based cell growth determination kit (Sigma-Aldrich) as previously described [3 (link)]. Relative cell survival was calculated as percentage relative to DMSO vehicle-treated controls. A dose-response curve was fitted to the data using the software KaleidaGraph 4.0 (Synergy), from which the half-maximal inhibitory concentration (IC50) was derived and represented as a mean of 4 independent experiments ± standard error of the mean (SEM).
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