Cell viability of primary hepatocytes and Kupffer cells after OA treatment was evaluated using the MTT kit (C0009S, Beyotime, Jiangsu, China) according to the manufacturer’s protocol. Briefly, primary hepatocytes and Kupffer cells were seeded into 96-well plates at 5 × 103 cells/well in quadruplicate and treated with 0, 0.1, 0.2, 0.4, 0.6, 0.8, 1.0, 1.2, and 1.5 mmol/L OA for 24 h. The MTT solution (10 μL) was then added to each well and the cells were cultured for 4 h in a humidified atmosphere of 95% air and 5% CO2 at 37°C. Subsequently, formazan solvent (100 μL) was added to each well and the cells were cultured for another 4 h until all the formazan dissolved. A microplate reader (SpectraMax i3; Molecular Devices LLC., San Jose, CA, United States) was used to measure the absorbance at 570 nm.
Mtt kit
Manufactured by Beyotime
Sourced in China
The MTT kit is a laboratory equipment used for cell viability and proliferation assays. It contains the necessary reagents and protocols to quantify viable cells in a simple and effective manner.
Automatically generated - may contain errors
Lab products found in correlation
Microplate reader, by Thermo Fisher Scientific (6 mentions)
Microplate reader, by Bio-Rad (4 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (3 mentions)
Facscanto flow cytometer, by BD (2 mentions)
Trizol extraction kit, by Thermo Fisher Scientific (2 mentions)
Sybr green pcr master mix, by Thermo Fisher Scientific (2 mentions)
Lipofectamine 2000 transfection kit, by Thermo Fisher Scientific (2 mentions)
Microplate reader, by Agilent Technologies (2 mentions)
Foetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Ripa lysis buffer, by Beyotime (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (2 mentions)
Nanodrop, by Thermo Fisher Scientific (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Spectramax i3, by Molecular Devices (1 mentions)
Transwell plate, by Merck Group (1 mentions)
Matrigel, by BD (1 mentions)
Abi steponeplus real time pcr system, by Thermo Fisher Scientific (1 mentions)
Annexin 5 pi apoptosis detection kit, by Thermo Fisher Scientific (1 mentions)
73 protocols using mtt kit
1
Cell Viability of Hepatocytes and Kupffer Cells after OA Exposure
2
Cell Proliferation and Migration Assays
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After 24 hours of transfection, cells were trypsinized and seeded into 96-well culture plate at a density of 10,000 cells/well in growth medium supplemented with 10% serum. The cell proliferation was measured at different time points (24, 48, and 72 hours) using the 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) kit (Beyotime Biotechnology, Nanjing, China), following the manufacturer's protocol. Cell migration was determined using 8 μm pore size 96-well minimum inhibitory concentration (MIC) Transwell plates (Millipore), via measuring the number of migratory cells under a microscope in 5 fields (100×), as previously described [21 (link)]. For the invasion assay, the MIC plates were initially coated with matrigel (BD Biosciences, Bedford, MA, USA), diluted in serum free medium and followed the same procedures as migration assay.
3
MTT Assay for IPEC-J2 Cell Viability
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The MTT kit was purchased from Beyotime Biotechnology (Shanghai, China) and used according to the manufacturer’s protocol. Briefly, IPEC-J2 cells were seeded in 96-well plates at a density of 4000 cells per well with 200 μL of complete culture medium. After being allowed to adhere and spread for 12 h, the cells were treated with 3 pmol of MIR167e-5p, NC, inhibitor, and i-NC for 0, 24, 48, and 72 h. MTT assays were performed by incubating the treated IPEC-J2 cells with 20 μL (5 mg/mL) of MTT labelling solution. After 4 h of incubation, IPEC-J2 were lysed with 150 μL of DMSO, and the purple formazan crystals were solubilized for detection at 570 nm.
4
Comparative Analysis of Retinal Cell Lines
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Y79 cells of human Rb cell line and ARPE-19 cells of human retinal epithelial cells (BNCC341293, BNCC337713) were purchased from BeNa Culture Collection. Lipofectamine™ 2000 Transfection Kit, ABI Stepone Plus Real-Time PCR System, Annexin V/PI Apoptosis Detection Kit, and Trizol Extraction Kit (Invitrogen, Carlsbed, CA, USA). SYBR Green PCR Master Mix (Applied Biosystems, Waltham, MA, USA). MTT kit (Beyotime Biotechnology Co., Ltd., Shanghai, China, product number: C0009). Transwell kit, 10% fetal bovine serum (FBS), FACSCanto flow cytometer (Becton Dickinson, Franklin Lakes, NJ, USA). DR5000 UV-visible spectrophotometer (BioRad, Hercules, USA). The design and synthesis of all the primer sequences were carried out by Sangong Bioengineering Co., Ltd., Shanghai, China.
5
In vitro Wound Healing Assay
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For in vitro wound-healing assays, wounds were made in confluent monolayers of TPC-1 cells using a pipette tip, and images were captured following co-culture with macrophages for 12 h. For cell proliferation assays, TPC-1 cells were co-cultured with macrophages at 37°C for 1, 3 and 5 days. MTT assays were performed at each time point using an MTT kit (Beyotime Institute of Biotechnology, Shanghai, China) according to the manufacturer's protocol.
6
MTT Assay for H2O2-Induced Cytotoxicity
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An MTT kit (Beyotime Institute of Biotechnology, Shanghai, China) was employed to measure cell viability. ARPE-19 cells were seeded into 96-well plates (2×103 cell/well) for 24 h. Cells were exposed to various concentrations of H2O2 (0, 10, 50, 100, 200, 400 and 800 µM) and GP (0, 5, 10, 30, 50 and 100 µM) for 24 h at 37°C, respectively. Other cells were transfected with NC and siNrf2 vectors as aforementioned. Then, cells were incubated with MTT solution for 4 h at 37°C. Subsequently, the supernatant was removed; dimethyl sulfoxide was then added to the cells and the optical density at 490 nm was evaluated using a microplate reader (SpectraMax iD3, Molecular Devices, LLC, Sunnyvale, CA, USA); 200 µM H2O2 was finally used to treat cells in the subsequent experiments.
7
Cell Viability and Morphometry Analysis
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Cell viability was detected using a commercial MTT kit (Beyotime Biotechnology, China) according to the manufacturer's instructions. An anti-actinin antibody (05-384, Millipore) was used to stain the cells according to our previous study [16 (link)]. Image-Pro Plus (version 6.0) was used to determine the surface areas of single cells.
8
Cell Viability Assay in BV2 Cells
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The BV2 cells (1 × 104/well) were seeded into 96-well plates for assessment of viability with an MTT kit (Beyotime, Shanghai, China).
9
MTT Cell Viability Assay Protocol
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VECs were seeded and added with MTT solution (5 mg/mL) for 4-h culture. Next, each well was added with 100 μL dimethyl sulfoxide for 10-min shaking. A microplate reader was used for measurement of optical density (OD) value at 490 nm. This assay was performed based on MTT kit instructions (Beyotime Institute of Biotechnology, Shanghai, China).
10
Cell Viability Quantification by MTT Assay
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MTT kit (Beyotime, Shanghai, China) was utilized to detect the cell viability. In brief, cells were cultured for 48 hours at a density of 5000 cells per well. After that, each well was incubated for 4 hours with 10 μL MTT solution, and then incubated with 100 μL Formazan dissolving solution until the formazan precipitate was completely dissolved through the observation under an optical microscope. A spectrophotometer (Unico Instrument Co., Ltd., Shanghai, China) was adopted to measure the optical density (OD) value at 570 nm.
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