Cell viability was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay kit (Boehringer Mannheim GmbH, Mannheim, Germany) as per the manufacturer’s protocols. In order to determine the cytotoxicity of type II collagen extract, cells were treated with 25, 50, 100, and 200 μg/mL for 24 hr. Cultures of the control group were left untreated. Ten μL of the MTT labeling reagent was added to each well, and the plates were incubated for 4 hr. Solubilization solution of 100 μL was then added to each well, and the cells were incubated for another 12 hr. The absorbance was then measured with a microtiter plate reader absorbance was then measured with a microtiter plate reader (Bio-Tek, Winooski, VT, USA) at a test wavelength of 195 nm and a reference wavelength of 690 nm. Optical density (O.D.) was calculated as the difference between the absorbance at the reference wavelength and that at the test wavelength. Percent viability was calculated as (O.D. of drug-treated sample/control O.D.) ×100.
Mtt assay kit
Manufactured by Roche
Sourced in Germany, United States
The MTT assay kit is a colorimetric assay used to measure cell metabolic activity. It utilizes the yellow tetrazolium salt MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide), which is reduced by metabolically active cells to form purple formazan crystals. The amount of formazan produced is directly proportional to the number of viable cells, and can be quantified by spectrophotometric analysis.
Automatically generated - may contain errors
Lab products found in correlation
Matrigel invasion chamber, by BD (3 mentions)
Prism 6, by GraphPad (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Lamin b, by Abcam (2 mentions)
Tumor necrosis factor (tnf), by Thermo Fisher Scientific (2 mentions)
M csf, by Thermo Fisher Scientific (2 mentions)
Phospho erk, by Cell Signaling Technology (2 mentions)
Blimp1, by Cell Signaling Technology (2 mentions)
Srankl, by Thermo Fisher Scientific (2 mentions)
I bet, by GlaxoSmithKline (2 mentions)
C myc and p38, by Santa Cruz Biotechnology (2 mentions)
Nfatc1, by BD (2 mentions)
Microtiter plate reader, by Agilent Technologies (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
Abt 199, by Merck Group (1 mentions)
Microplate reader, by Awareness Technology (1 mentions)
Ficoll paque density gradient, by Cedarlane (1 mentions)
5 μm pore size inserts, by Corning (1 mentions)
Macs cell separation technology, by Miltenyi Biotec (1 mentions)
Microplate reader, by Bio-Rad (1 mentions)
49 protocols using mtt assay kit
1
Cell Viability Evaluation by MTT Assay
2
Rocuronium Bromide and LPS Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Rocuronium bromide was obtained from Korea Organon (Seoul, Korea). LPS was obtained from Sigma Chemical Co. (St. Louis, MO, USA). MTT assay kit was purchased from Boehringer Mannheim (Mannheim, Germany).
3
MTT Assay for Cell Viability
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell viability was determined using an MTT assay kit (Roche Diagnosis, Indianapolis, IN) according to the manufacturer's protocol. Briefly, stable cell lines were plated in 96-well plates (2, 000 cells/well) and cell viability was determined after 48 h.
4
Cytotoxicity Assay of Paclitaxel in Transfected Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Transfected cells were cultured for 24 h and treated with paclitaxel at the desired concentration for each cell line (Table 1 ). DMSO served as vehicle control. The treated cells were incubated for 48 h and a cytotoxicity assay was performed using an MTT assay kit (Roche, 11465007001) according to the manufacturer protocol. Briefly, 10 μl MTT (5 mg/ml) was added to each well and allowed to form formazan crystals for four hours in the incubator. In total, 100 μl of solubilization solution was added to each well and incubated overnight in the incubator in a humidified atmosphere. The next day, complete solubilization of the purple formazan crystals was confirmed, and then the absorbance values were determined using a microplate reader (BMG FLUOstar Omega) at 590 nm. The experiments were repeated twice, and data are represented as mean ± SD from three technical replicas. No blinding was done.
5
Cytotoxic Effects of miRNA-16-1 and ABT-199 on CLL Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cytotoxic effects of miRNA-16-1 and ABT-199 (Sigma- Aldrich, USA) on
CLL cells was evaluated using 3-(4 (link), 5 (link)-Dimethylthiazol-2-yl)-2, 5-Diphenyltetrazolium
Bromide (MTT) assay (25 (link)). The experiment was divided into eight groups: ABT-199,
miRNA-16-1 mimics, NC miRNA, miRNA-16-1 mimics and
ABT-199, NC miRNA and ABT-199, miRNA blank control, ABT-199 blank control and combination
blank control. Briefly, the cells were cultivated in 96-well tissue plates at a density of
5×104 cells per well, and then transfected with miRNAs. Six hours later, the
cells were treated with different concentrations of ABT-199 (0, 0.05, 0.1, 0.2, 0.4, 0.8,
1.6 and 3.2 µM). After 24 and 48 hours of incubation, the cytotoxicity was determined
using a MTT assay kit (Roche Diagnostics GmbH, Mannheim, Germany) according to the
manufacturer’s instructions. The absorbance (A) was measured spectrophotometrically at 570
nm with a microplate reader (Awareness Technology, Palm City, FL, USA). Half-maximal
inhibitory concentration (IC50) (drug concentration that reduced 50% survival
rate) value of the ABT-199, alone or in combination with miRNA, was calculated with Prism
6.01 software (GraphPad Software Inc., San Diego, CA, USA). In the next experiments, the
IC50 doses of fludarabine were used.
CLL cells was evaluated using 3-(4 (link), 5 (link)-Dimethylthiazol-2-yl)-2, 5-Diphenyltetrazolium
Bromide (MTT) assay (25 (link)). The experiment was divided into eight groups: ABT-199,
miRNA-16-1 mimics, NC miRNA, miRNA-16-1 mimics and
ABT-199, NC miRNA and ABT-199, miRNA blank control, ABT-199 blank control and combination
blank control. Briefly, the cells were cultivated in 96-well tissue plates at a density of
5×104 cells per well, and then transfected with miRNAs. Six hours later, the
cells were treated with different concentrations of ABT-199 (0, 0.05, 0.1, 0.2, 0.4, 0.8,
1.6 and 3.2 µM). After 24 and 48 hours of incubation, the cytotoxicity was determined
using a MTT assay kit (Roche Diagnostics GmbH, Mannheim, Germany) according to the
manufacturer’s instructions. The absorbance (A) was measured spectrophotometrically at 570
nm with a microplate reader (Awareness Technology, Palm City, FL, USA). Half-maximal
inhibitory concentration (IC50) (drug concentration that reduced 50% survival
rate) value of the ABT-199, alone or in combination with miRNA, was calculated with Prism
6.01 software (GraphPad Software Inc., San Diego, CA, USA). In the next experiments, the
IC50 doses of fludarabine were used.
6
Macropinocytosis, Slit2, and Cell Viability
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
DLD-1 and PANC-1 cells were seeded (104 cells per well) in a 96-well plate and grown in DMEM without l -Glut supplemented with 0.5 mM Glut (physiological levels), 0.2 mM Glut (low Glut condition), and 2% albumin, when indicated, for 6 days. The macropinocytosis inhibitor, EIPA (25 µM), was added to the medium for the last 2 days only, where indicated. In some experiments, 30 nM NSlit2 or Slit2Δ2 was added and the medium was changed every 24 h, for 6 days. At the end of all treatments, cell viability was assessed using a MTT assay kit (Roche Diagnostics purchased from Sigma-Aldrich, Oakville, ON, Canada).
7
Cell Viability Assay for Drug Screening
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The
duplicate set of
cells were treated with same drug concentrations but left uninfected.
After 48 h of incubation at 37 °C, the cells were analyzed for
viability using MTT assay kit (Roche). The viability was calculated
as a percentage relative to DMSO control in triplicates.
duplicate set of
cells were treated with same drug concentrations but left uninfected.
After 48 h of incubation at 37 °C, the cells were analyzed for
viability using MTT assay kit (Roche). The viability was calculated
as a percentage relative to DMSO control in triplicates.
8
Evaluating T-cell Chemotaxis in Lymphoma Microenvironment
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Peripheral blood mononuclear cells were separated from the peripheral blood of two healthy donors using a Ficoll‐Paque density gradient (Cedarlane), and total T cells were collected by negative selection using MACS Cell Separation Technology (Miltenyi Biotec). Chemotaxis of T cells was evaluated using 24‐well migration chambers with 5‐μm‐pore‐size inserts (Corning). Lower chambers were filled with 600 μl of the supernatants of lymphoma cells cultured with the indicated concentrations of tazemetostat or DMSO for 4 days (FL318) or 3 days (primary lymphoma cells), and 2 × 106 T cells were plated in the upper chambers and incubated at 37°C for 12 hours. In blocking experiments using the anti‐CCR4 antibody, T cells were incubated for 1 hour with different concentrations of anti‐CCR4 antibody (Cayman Chemical) before the cell migration assay. T cells that migrated from the upper to the lower chambers were collected, and the cell numbers were calculated using an MTT assay kit (Roche Diagnostics), and the T‐cell subpopulation was analyzed by flow cytometry after migration.
9
Cell Viability Monitoring via MTT Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were seeded in a 96-well plate with 1 × 104 cells/well. Cell viability was assessed every 24 hours using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay kit (Roche Applied Science, Mannheim, Germany) according to the manufacture’s instruction. Experiments were repeated 3 times independently.
10
Angiogenin Regulates HUVEC Proliferation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HUVECs were seeded in six-well plate at 20% confluency in culture medium and incubated overnight at 37 °C. Upon treatment, cell culture medium was changed into vascular cell basal medium containing 1% FBS, and rhANG (1 mg/mL, kindly provided by Dr. Guo-Fu Hu) or individual CM media prepared as described above (1:1 with vascular cell basal medium containing 1% FBS) was added and cultured for 24 h. Cell proliferation was assayed using MTT assay kit (Roche, 11465007001) according to the manufacturer’s instruction. MTT assays were performed in quadruplicates each time and repeated three times.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!