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Thiazolyl blue

Thiazolyl blue, also known as 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), is a widely used tetrazolium-based colorimetric assay for assessing cell metabolic activity and viability.
It is a versatile reagent employed in a variety of biomedical and pharmaceutical research applications, including cell proliferation, cytotoxicity, and cell-based assays.
PubCompare.ai can enhance your Thiazolyl blue research by locating optimal protocols from scientific literature, pre-prints, and patents, helping you identify the most reproducible and accurate methods and boosting your research productivity.
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Most cited protocols related to «Thiazolyl blue»

1
Dual-Luciferase Assay for microRNA Targeting
Cited 14 times
A full length of the human PLAU 3′-untranslated region (932 bp) and partial length of HIC2 3′-UTR (1131 bp, 1862-2992 from a 4719-bp full length) with the miR-193a-3p targeting sequence were cloned at the downstream of the firefly luciferase gene in pGL3 (Invitrogen) to construct pGL3-luc-PLAU and pGL3-luc-HIC2, respectively. All the constructs were confirmed by restriction digestion.
Cells were seeded into 96-well plates at around 1 × 104 cells per well and transfected with a mixture of 50 ng pGL3-luc-PLAU or pGL3-luc-HIC2, 5 ng Renilla plus 5 pmol mimic or scramble control (NC) nucleotides, with the riboFECT CP transfection reagents according to the manufacturer's instruction. Both firefly and Renilla luciferase activities were measured 18 h after transfection by the Dual-Luciferase Reporter Assay System (Promega) using a Promega GloMax 20/20 luminometer. The relative firefly luciferase activities were normalized with the Renilla luciferase activities as a for transfection efficiency.
The pathway luciferase reporter constructs (Supplementary Figure S2A): (1) the negative control construct: the firefly luciferase gene is under the control of the minimal promoter. (2) the pathway reporter construct: a tandem repeat of the cognate consensus motif that is recognized by each master transcription factor for the corresponding pathway was placed at the upstream of the minimal promoter in the construct 1. (3) the positive control construct: the firefly luciferase gene is under the control of CMV promoter and (4) the internal control construct. The firefly luciferase gene in construct 3 was replaced with the Renilla luciferase gene. The analysis was carried out according to the manufacturer's instruction (Qiagen).42 (link) Briefly, the cells were transfected in triplet with each firefly luciferase reporter construct in combination with the Renilla luciferase construct using ribo FECT CP transfection reagent, and both luciferase activities in cell extracts at 24 h after transfection were measured by a Promega Dual-Luciferase Reporter assay (Promega) using a Promega GloMax 20/20 luminometer.43 Firefly luciferase activities from each set were normalized to the activity of Renilla luciferase to control the inter-transfection bias. The relative luciferase activities (luciferase unit) of the pathway reporter over the negative control in the transfected cells were calculated as a measurement of the pathway activity.
Chemotherapeutics: All the chemotherapeutic drugs used are of the clinic grade44 (link), 45 (link) (NCI Dictionary of Cancer Terms, http://www.cancer.gov/dictionary), Pi: Pirarubicin hydrochloride (Wanle, Shenzhen, China); Pa: Paclitaxel (Shuanglu, Beijing, China); Ad: Adriamycin (Haizheng, Zhejiang, China); EH: Epirubicin hydrochloride (Haizheng, Zhejiang, China).
Chemoresistance profiling (IC50 measurements): Cells in the logarithmic phase of growth were seeded in triplicate in 96-well plates at the density of 0.5 × 104/well and treated with four-fold serially diluted drugs for 72 h. Cell survival was then measured by a thiazolyl blue tetrazolium bromide (MTT, 490 nm reading)-based cell proliferation assay.46 (link) Both the linear regression parameters and the IC50 (the concentration of drug required for 50% of cells to be killed) with the no-drug control as the reference were calculated. The relative chemoresistance was presented as the fold for each of the cell line over the lowest IC50.
Lv L., Deng H., Li Y., Zhang C., Liu X., Liu Q., Zhang D., Wang L., Pu Y., Zhang H., He Y., Wang Y., Yu Y., Yu T, & Zhu J. (2014). The DNA methylation-regulated miR-193a-3p dictates the multi-chemoresistance of bladder cancer via repression of SRSF2/PLAU/HIC2 expression. Cell Death & Disease, 5(9), e1402-.
Publication 2014
2
Quantification of Cellular NADPH, NADP, GSH, and ATP
Cited 8 times
The intracellular levels of NADPH and total NADP (NADPH+NADP+) were measured with previously described enzymatic cycling methods, with modifications32 (link),33 (link). In brief, 1.8 × 106 cells were plated in 10-cm dishes; on the next day, the cells were lysed in 400 µl of extraction buffer (20 mM nicotinamide, 20 mM NaHCO3, 100 mM Na2CO3) and centrifuged. For NADPH extraction, 150 µl of the supernatant was incubated at 60 °C for 30 min. Next, 160 µl of NADP-cycling buffer (100 mM Tris-HCl pH 8.0, 0.5 mM thiazolyl blue, 2 mM phenazine ethosulfate, 5 mM EDTA) containing 1.3U of G6PD was added to a 96-well plate containing 20 µl of the cell extract. After a 1-min incubation in the dark at 30 °C, 20 µl of 10 mM glucose 6-phosphate (G6P) was added to the mixture, and the change in absorbance at 570 nm was measured every 30 s for 4 min at 30 °C with a microplate reader. The concentration of NADP+ was calculated by subtracting[NADPH] from [total NADP]. The intracellular levels of GSH and total glutathione (GSSG + GSH) were measured with the use of enzymatic cycling methods, as described previously34 . The intracellular level of ATP was measured with an ATPlite assay kit (Perkin Elmer).
Jeon S.M., Chandel N.S, & Hay N. (2012). AMPK regulates NADPH homeostasis to promote tumour cell survival during energy stress. Nature, 485(7400), 661-665.
Publication 2012
5-ethylphenazine Bicarbonate, Sodium Biological Assay Buffers Cell Extracts Cells Edetic Acid Enzymes Glucose-6-Phosphate Glucosephosphate Dehydrogenase Glutathione Disulfide Hyperostosis, Diffuse Idiopathic Skeletal NADP Niacinamide Protoplasm thiazolyl blue Tromethamine
3
MTT Assay for Cytotoxicity Evaluation
Cited 5 times
Cell cytotoxicity was determined with the MTT based-In Vitro Toxicology Assay Kit. Cancer cells were treated with cottonseed extracts and incubated at 37 °C, 5% CO2 for 2 and 24 h. The cell media were added with 50 µl of MTT assay reagent (thiazolyl blue tetrazolium bromide) and incubated at 37 °C, 5% CO2 for 2 h before adding 500 µl MTT solubilization solution to each well, shaken at room temperature overnight. The color density in the wells was recorded by Epoch microplate spectrophotometer at A570 after diluting 8 times.
Cao H., Sethumadhavan K, & Bland J.M. (2018). Isolation of Cottonseed Extracts That Affect Human Cancer Cell Growth. Scientific Reports, 8, 10458.
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Publication 2018
Biological Assay Cells Cytotoxin EPOCH protocol Malignant Neoplasms thiazolyl blue tetrazolium bromide
4
miR-485-5p Regulates Cell Proliferation
Cited 5 times
Twenty-four hours after transfection with miR-485-5p inhibitor, miR-485-5p mimics, miRNA mimic negative controls (NC miR-mimics) and miRNA inhibitor negative controls, NCI-H446 and NCI-H1688 cells were harvested and sub-cultured in 96-well plates. Cell proliferation was assessed using thiazolyl blue tetrazolium bromide assay (MTT; Amresco, Radnor, PA, USA) according to the manufacturer’s instructions. Briefly, MTS reagent (20 µl) was added to each well and incubated at 37°C for 4 h. Then, the reagent was removed and dimethyl sulfoxide (150 µl) was added to each well. Absorbance at 492 nm was measured using the FLx800 fluorescence microplate reader (BioTek, Winooski, VT, USA). The experiment was performed in triplicate and repeated three times. The data are expressed as means ± standard error of the mean (SEM).
Gao F., Wu H., Wang R., Guo Y., Zhang Z., Wang T., Zhang G., Liu C, & Liu J. (2019). MicroRNA-485-5p suppresses the proliferation, migration and invasion of small cell lung cancer cells by targeting flotillin-2. Bioengineered, 10(1), 1-12.
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Publication 2019
Biological Assay Cell Proliferation Cells Fluorescence MicroRNAs Sulfoxide, Dimethyl thiazolyl blue tetrazolium bromide Transfection
5
Propagation and Titration of RSV
Cited 5 times
Human RSV strain A2 (ATCC, Manassas, VA; #VR-1540) was infected at a multiplicity of 0.1 into Hep2 cells. The virus was allowed to grow for 5 days at 37°C in a 5% CO2 atmosphere. The infected Hep2 monolayers were collected and the virus was released by sonication. Cell debris was removed by centrifugation at 2500 g for 5 minutes at 4°C. Virus was collected by centrifuging the supernatant for 2 hours at 22000×g at 4°C. Virus were suspended in culture media and snap frozen and maintained at −80°C. Infectious virus titers were determined on Hep2 cells by performing serial dilutions of the RSV stocks and counting infected cells stained for indirect immunofluorescence with an RSV F-specific monoclonal antibody (Abcam, Cambridge, MA). Additionally, plaque assays were performed as previously described [25] on Hep2 cells using methyl cellulose overlay media (R&D Systems) and staining with 0.5 mg/ml thiazolyl blue tetrazolium bromide (MTT; Sigma Aldrich) solution in PBS for 3 hours at 37°C. Non-infected Hep2 cell cultures were processed in the same manner as RSV infected cells and the resulting sample collection was used as a mock control.
Foronjy R.F., Dabo A.J., Taggart C.C., Weldon S, & Geraghty P. (2014). Respiratory Syncytial Virus Infections Enhance Cigarette Smoke Induced COPD in Mice. PLoS ONE, 9(2), e90567.
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Publication 2014
Atmosphere Biological Assay Cell Culture Techniques Cells Centrifugation Culture Media Dental Plaque Freezing Human respiratory syncytial virus Indirect Immunofluorescence Infection Methylcellulose Monoclonal Antibodies Specimen Collection Strains Technique, Dilution thiazolyl blue tetrazolium bromide Virus

Most recents protocols related to «Thiazolyl blue»

1
MTT Assay for Cell Viability
Cell metabolic activity and cell viability were determined using a Thiazolyl Blue Tetrazolium Bromide (MTT) (M5655, Sigma, Poole, UK) assay, as described previously [51 (link)]. SHSY-5Y cells were seeded at 3 × 104 cells/well in 96-well plates with growth medium (10% FBS). After 24 h, undifferentiated cells were exposed to ethanol (0–200 mM) diluted in growth media (10% FBS). Differentiated cells were prepared as described above and then treated with ethanol (0–200 mM) diluted in differentiation medium supplemented with 20 ng/mL BDNF. After incubation, spent medium was removed and then replaced with medium containing 10% 5 mg/mL MTT and incubated for 4 h. Plate wells which only received 10% MTT and respective growth medium served as background controls. The generated formazan crystals were suspended in a 1:1 dimethyl sulphoxide (DMSO, D8418, Sigma, Poole, UK)–isopropanol (279544, Sigma, Poole, UK) solution. The absorbance of wells was then read at 570 nm using a spectrophotometer (Multiskan Spectrum, Thermo Electron Corporation, Waltham, MA, USA). An average value was calculated from experiments performed in triplicate after the subtraction of blank (negative control) values. Cell viability was expressed as a percentage of survival compared with that from mock-treated cells. The inhibitor concentrations producing 50% loss of viability of cells (IC50 values) were obtained from the concentration–response curves and expressed as mean ± standard deviation (SD).
Mudyanselage A.W., Wijamunige B.C., Kocoń A., Turner R., McLean D., Morentin B., Callado L.F, & Carter W.G. (2024). Alcohol Triggers the Accumulation of Oxidatively Damaged Proteins in Neuronal Cells and Tissues. Antioxidants, 13(5), 580.
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Publication 2024
2
Cell Viability Assay Protocol
Cells were seeded into microtiter plates (Life Sciences, Corning, NY, USA) at a concentration of 5 × 103 cells/100 µL/well in full medium and allowed to attach overnight, followed by the incubation in fresh full medium with or without the test substances (at concentrations of 0.005, 0.01, 0.025, 0.05, 0.1, 0.25, 0.5, and 1 µg/mL for nintedanib and 1, 10, 50, 125, 250, 500, 750, and 1000 µg/mL for the other 3 substances) for 48 hours. The stock solution of MTT (5 mg/mL in PBS; Gibco by Lifetechnologies, Carlsbad, CA, USA) was added at a dilution of 1:5 (v/v) and the cells were incubated further for 3.5 hours.64 (link),65 (link) Measurement of absorbance (UV/visual absorption spectroscopy in optical density [OD]) was performed using a spectrophotometer (Molecular Devices GmbH, Biberach an der Riss, Germany) at 570 nm64 after dissolving the MTT crystals with DMSO.66 (link) Negative controls were performed by measuring wells without cells but with the addition of MTT solution to prevent false positive results.
Pawlik V.E., Sonntag S.R., Grisanti S., Tura A., Kakkassery V, & Ranjbar M. (2024). Impact of Nintedanib and Anti-Angiogenic Agents on Uveal Melanoma Cell Behavior. Investigative Ophthalmology & Visual Science, 65(2), 30.
Publication 2024
3
MTT Cell Viability Assay
The MTT assay was carried out in accordance with the supplier’s guidelines (Sigma-Aldrich, cat#M5655, St. Louis, MO, USA). Briefly, the culture medium was cautiously removed from each plated well following each treatment. Each well was then filled with 50 µL of serum-free medium and 50 µL of MTT® solution, and the mixture was incubated for three hours at 37 °C. Each well received 150 µL of MTT solvent after incubation. For fifteen minutes, the foil-wrapped plate was shaken on an orbital shaker. Sometimes, the liquid needed to be pipetted in order to completely dissolve the MTT formazan. Within an hour, plate absorbance at OD = 590 nm was measured. In three separate experiments, the evaluation was conducted again. The percentage of cell viability was calculated using the following equation: % Viability = Mean OD(untreated sample) − Mean OD(treated sample)/Mean OD(untreated sample) × 100. Samples were reported in comparison to the negative control (untreated wild-type ChLN cells) considered to have 100% viability.
Gomez-Sequeda N., Jimenez-Del-Rio M, & Velez-Pardo C. (2024). Combination of Tramiprosate, Curcumin, and SP600125 Reduces the Neuropathological Phenotype in Familial Alzheimer Disease PSEN1 I416T Cholinergic-like Neurons. International Journal of Molecular Sciences, 25(9), 4925.
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Publication 2024
4
Evaluating ADP Cytotoxicity by MTT and Flow
The cytotoxicity of different ADPs was evaluated by MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide, thiazolyl blue tetrazolium bromide) assay and flow cytometry. The cells were treated as above mentioned. Subsequently, Annexin V fluorescein isothiocyanate/propidium iodide double-staining kit was used to assess the cell apoptosis and the MTT was performed to assess the cell viability.
Zhu J., Peng L., Jehan S., Wang H., Chen X., Zhao S, & Zhou W. (2024). Activable Photodynamic DNA Probe with an “AND” Logic Gate for Precision Skin Cancer Therapy. Research, 7, 0295.
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Publication 2024
5
MTT-Based Cell Viability Assay
Cell viability was measured by a colorimetric assay using MTT (Thiazolyl Blue Tetrazolium Bromide, Sigma-Aldrich, St. Louis, MO, USA) as described before. The data is presented as relative changes in percent compared to untreated controls [26 (link)].
Pan Y., Xin W., Wei W., Tatenhorst L., Graf I., Popa-Wagner A., Gerner S.T., Huber S.E., Kilic E., Hermann D.M., Bähr M., Huttner H.B, & Doeppner T.R. (2024). Knockdown of NEAT1 prevents post-stroke lipid droplet agglomeration in microglia by regulating autophagy. Cellular and Molecular Life Sciences: CMLS, 81(1), 30.
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Publication 2024

Top products related to «Thiazolyl blue»

1
Thiazolyl blue tetrazolium bromide by Merck Group
Sourced in United States, Germany, United Kingdom, Italy, Spain, Macao, Sao Tome and Principe, Ireland, India, France, Japan, Poland
Thiazolyl blue tetrazolium bromide is a colorimetric reagent commonly used in cell viability assays. It is a yellow tetrazolium salt that is reduced to a purple formazan product by viable cells, enabling the quantification of cell proliferation or cytotoxicity.
2
Dimethyl sulfoxide (dmso) by Merck Group
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DMSO is a versatile organic solvent commonly used in laboratory settings. It has a high boiling point, low viscosity, and the ability to dissolve a wide range of polar and non-polar compounds. DMSO's core function is as a solvent, allowing for the effective dissolution and handling of various chemical substances during research and experimentation.
3
Methyl thiazolyl tetrazolium (mtt) by Merck Group
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MTT is a colorimetric assay used to measure cell metabolic activity. It is a lab equipment product developed by Merck Group. MTT is a tetrazolium dye that is reduced by metabolically active cells, producing a colored formazan product that can be quantified spectrophotometrically.
4
Thiazolyl blue tetrazolium bromide mtt by Merck Group
Sourced in United States, Germany, United Kingdom, Japan, Italy, China, Macao, Sao Tome and Principe, France
Thiazolyl blue tetrazolium bromide (MTT) is a yellow tetrazolium salt that can be reduced to a purple formazan product by dehydrogenase enzymes, primarily those found in the mitochondria of living cells. This reduction process is used as a widely accepted method for assessing cell viability and proliferation in various cell-based assays.
5
Fetal bovine serum (fbs) by Thermo Fisher Scientific
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Fetal Bovine Serum (FBS) is a cell culture supplement derived from the blood of bovine fetuses. FBS provides a source of proteins, growth factors, and other components that support the growth and maintenance of various cell types in in vitro cell culture applications.
6
Penicillin streptomycin by Thermo Fisher Scientific
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Penicillin/streptomycin is a commonly used antibiotic solution for cell culture applications. It contains a combination of penicillin and streptomycin, which are broad-spectrum antibiotics that inhibit the growth of both Gram-positive and Gram-negative bacteria.
7
Propidium iodide by Merck Group
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Propidium iodide is a fluorescent dye commonly used in molecular biology and flow cytometry applications. It binds to DNA and is used to stain cell nuclei, allowing for the identification and quantification of cells in various stages of the cell cycle.
8
Fetal bovine serum (fbs) by Merck Group
Sourced in United States, Germany, United Kingdom, Japan, Italy, China, Macao, Switzerland, France, Canada, Sao Tome and Principe, Spain, Australia, Ireland, Poland, Belgium, Denmark, India, Sweden, Israel, Austria, Brazil, Czechia, Netherlands, Portugal, Norway, Holy See (Vatican City State), New Zealand, Hungary, Senegal, Argentina, Thailand, Singapore, Ukraine, Mexico
FBS, or Fetal Bovine Serum, is a commonly used cell culture supplement. It is derived from the blood of bovine fetuses and provides essential growth factors, hormones, and other nutrients to support the growth and proliferation of a wide range of cell types in vitro.
9
Trypsin edta by Thermo Fisher Scientific
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Trypsin-EDTA is a solution used in cell culture applications to dissociate adherent cells from their growth surface. It contains the proteolytic enzyme trypsin and the chelating agent EDTA, which work together to break down the cellular adhesions and allow the cells to be harvested and passaged.
10
Phosphate buffered saline (pbs) by Merck Group
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PBS (Phosphate-Buffered Saline) is a widely used buffer solution in biological and medical research. It is a balanced salt solution that maintains a stable pH and osmotic pressure, making it suitable for a variety of applications. PBS is primarily used for washing, diluting, and suspending cells and biological samples.

More about "Thiazolyl blue"

Thiazolyl blue, also known as 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), is a widely used tetrazolium-based colorimetric assay for assessing cell metabolic activity and viability.
This versatile reagent is employed in a variety of biomedical and pharmaceutical research applications, including cell proliferation, cytotoxicity, and cell-based assays.
The MTT assay works by measuring the reduction of the yellow tetrazolium salt (MTT) to purple formazan crystals by metabolically active cells.
This reduction is facilitated by the enzyme succinate dehydrogenase, which is present in the mitochondria of living cells.
The amount of formazan produced is directly proportional to the number of viable cells, making this a useful method for quantifying cell viability and proliferation.
Researchers often use MTT in conjunction with other cell culture components, such as DMSO (dimethyl sulfoxide) to solubilize the formazan crystals, and FBS (fetal bovine serum), penicillin/streptomycin, and trypsin-EDTA for maintaining and passaging cell lines.
Additionally, propidium iodide, a fluorescent dye, can be used in combination with MTT to distinguish between viable and non-viable cells.
PubCompare.ai can enhance your Thiazolyl blue (MTT) research by locating optimal protocols from scientific literature, pre-prints, and patents.
Our AI-driven comparisons help you identify the most reproducible and accurate methods, boosting your research productivity.
Experience the power of PubCompare.ai today!