Thiazolyl blue tetrazolium bromide

Manufactured by Thermo Fisher Scientific

Sourced in United States, United Kingdom

Thiazolyl blue tetrazolium bromide is a colorimetric reagent commonly used in cell viability assays. It functions as a redox indicator, changing color upon reduction by metabolically active cells.

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Lab products found in correlation

Lsm 880, by Zeiss (2 mentions) Hoechst 33342, by Thermo Fisher Scientific (2 mentions) Dimethyl sulfoxide (dmso), by Merck Group (2 mentions) Glyceraldehyde 3 phosphate dehydrogenase, by Cell Signaling Technology (1 mentions) Nf κb p65, by Cell Signaling Technology (1 mentions) C fos, by Cell Signaling Technology (1 mentions) Phospho nf κb p65, by Cell Signaling Technology (1 mentions) Phospho iκbα, by Cell Signaling Technology (1 mentions) C jun, by Cell Signaling Technology (1 mentions) Agarose, by Merck Group (1 mentions) 2 7 dichlorofluorescein diacetate, by Merck Group (1 mentions) Rhodamine 6g, by Merck Group (1 mentions) Dulbecco s phosphate bufferedsolution pbs, by Merck Group (1 mentions) Dulbecco s modified eagle s medium dmem, by Merck Group (1 mentions) Coumarin 6, by Merck Group (1 mentions) Indocyanine green icg, by Chem-Impex (1 mentions) Polystyrene standards, by Merck Group (1 mentions) Phosphotungstic acid pta, by Electron Microscopy Sciences (1 mentions) U87 mg cell lines, by RIKEN BioResource Center (1 mentions) Dulbecco s modified eagle medium dmem powder, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Thiazolyl Blue tetrazolium bromide (MTT) (14 citations)

18 protocols using thiazolyl blue tetrazolium bromide

Lipopolysaccharide-Induced Inflammation Signaling

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LPS from E. coli O111:B4 (LPS) and ammonium pyrrolidine
dithiocarbamate (APDC) were purchased from Sigma-Aldrich (St. Louis, MO, USA).
Thiazolyl Blue tetrazolium bromide (MTT) was obtained from Alfa Aesar
(Haverhill, MA, USA). Antibodies against cyclooxygenase-2 (COX-2), phospho-p38
MAPK, p38 MAPK, phospho-extracellular signal-regulated kinase 1/2 (ERK1/2),
ERK1/2, phospho-c-Jun N-terminal kinase (JNK), JNK, c-Fos, c-Jun,
phospho-IκBα, IκBα, phospho-p65 NF-κB, p65
NF-κB, phospho-AMP-activated protein kinase (AMPK), AMPK,
phospho-acetyl-CoA carboxylase (ACC), ACC, and glyceraldehyde-3-phosphate
dehydrogenase were obtained from Cell Signaling Technology (Danvers, MA, USA).
The anti-inducible nitric oxide synthase (iNOS) antibody was obtained from
GeneTex (Irvine, CA, USA).

Bae W.Y., Jung W.H., Shin S.L., Kwon S., Sohn M, & Kim T.R. (2022). Investigation of Immunostimulatory Effects of Heat-Treated Lactiplantibacillus plantarum LM1004 and Its Underlying Molecular Mechanism. Food Science of Animal Resources, 42(6), 1031-1045.

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Polymer-based Drug Delivery System

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Acid-terminated poly(d,l-lactic-co-glycolic acid) (PLGA)
(RG502H 7000–17 000 Da, viscosity: 0.16–0.24
dL/g, 0.1% (w/v) in chloroform), poly(allylamine hydrochloride) (PAH),
poly(diallyldimethylammonium chloride) (PDDA), (3-aminopropyl)trimethoxysilane
(APTMS), poly(sodium 4-styrenesulfonate) (PSS), agarose, poly(vinyl
alcohol) (PVA), coumarin-6, rhodamine 6G, Dulbecco’s phosphate-buffered
solution (PBS), 2′,7′-dichlorofluorescein diacetate
(DCFH-DA), and Dulbecco’s modified Eagle’s medium (DMEM)
were purchased from Sigma-Aldrich (MO). Tween 80 was obtained from
Fisher BioReagent (NJ). Solutol (Kolliphor HS 15) and erythromycin
were purchased from Glentham (U.K.), and indocyanine green (ICG) was
obtained from Chem-Impex (IL). Thiazolyl blue tetrazolium bromide
(MTT) was purchased from Alfa Aesar (Lancashire, U.K.).

Fluksman A., Lafuente A., Braunstein R., Steinberg E., Friedman N., Yekhin Z., Roca A.G., Nogues J., Hazan R., Sepulveda B, & Benny O. (2023). Modular Drug-Loaded Nanocapsules with Metal Dome Layers as a Platform for Obtaining Synergistic Therapeutic Biological Activities. ACS Applied Materials & Interfaces, 15(43), 50330-50343.

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ZDF VSMC Viability Assay with CA

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ZDF VSMC (5 × 10³ cells/well) were seeded in 96-well plates. Synchronized ZDF VSMC were treated with a range [6–1600 μM] of CA for 24 h. ZDF VSMC were treated with 0.4 mg/mL Thiazolyl Blue tetrazolium bromide (L11939; Alfa Aesar, Haverhill, MA) diluted in low glucose FBS-supplemented complete media for 4 h at 37 °C. After treatment, media was removed and plates were left to air-dry overnight. Formazan crystals were resuspended in DMSO and absorbance was measured at 560 nm with background at 670 nm on a Cytation 5 plate reader.

Buglak N.E., Jiang W, & Bahnson E.S. (2018). Cinnamic aldehyde inhibits vascular smooth muscle cell proliferation and neointimal hyperplasia in Zucker Diabetic Fatty rats. Redox Biology, 19, 166-178.

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Polymer-based Drug Delivery System Synthesis

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Poly(D,L-lactide-co-glycolide) 50:50 was from Evonik Industries (Birmingham, AL, USA). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC), N-ethyldiisopropylamine (N,N-Diisopropylethlamine) (DIEA, 99%), and thiazolyl blue tetrazolium bromide (MTT, 98%) were from Alfa Aesar (Heysham, UK). N-hydroxysuccinimide (NHS, 98%) was from Acros Organics Co. Inc. (Fair Lawn, NJ, USA). T7-peptide (FITC-Asp-His-Ala-Ile-Tyr-Pro-Arg-His-OH) and R9-peptide (Arg-Arg-Arg-Arg-Arg-Arg-Arg-Arg-Arg-OH) were synthesized by Kelowna International Scientific Inc. (Taipei, Taiwan). Palbociclib (PBC, 98%) was from Hunan HuaTeng Pharmaceutical Co., Ltd. (Changsha City, Hunan, China). Polystyrene standards were from Sigma-Aldrich Co., Ltd. (St. Louis, MO, USA). Poly(vinyl alcohol) (PVA, 88% hydrolyzed) was from Acros Organics Co. Inc. (Fair Lawn, NJ, USA). Phosphotungstic acid (PTA) was from Electron Microscopy Sciences (Hatfield, PA, USA). bEnd.3 and U87-MG cell lines were from Bioresource Collection and Research Center (Hsinchu, Taiwan). Dulbecco’s Modified Eagle Medium (DMEM) powder was from Thermo Fisher Scientific Inc. (Grand Island, NY, USA).

Lo Y.C, & Lin W.J. (2023). Improve BBB Penetration and Cytotoxicity of Palbociclib in U87-MG Glioblastoma Cells Delivered by Dual Peptide Functionalized Nanoparticles. Pharmaceutics, 15(10), 2429.

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Functionalized Nanoparticle-Based Cancer Therapy

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1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC), N-ethyldiisopropylamine (N,N-diisopropylethlamine) (DIEA, 99%), and thiazolyl blue tetrazolium bromide (MTT, 98%) were from Alfa Aesar (Echo Chemical Co., Ltd., Heysham, UK). N-hydroxysuccinimide (NHS, 98%) and poly(vinyl alcohol) (PVA, 88% hydrolyzed, 20,000–30,000 g/mol) were from Acros Organics Co., Inc. (Fair Lawn, NJ, USA). Poly(d,l-lactide-co-glycolide) 50:50 (PLGA, ~52,000 g/mol) was from Evonik Industries (Birmingham, AL, USA). Maleimide poly(ethylene glycol) amine (Mal-PEG-amine, 5000 g/mol) was provided by Hunan Hua Teng Pharmaceutical Co., Ltd. (Merelbeke, Belgium). FITC-NHS (MW 473.4 g/mol) was from Thermo Fisher Scientific Inc. (Hudson, NH, USA). FITC-Cys-T7 peptide (1498.71 g/mol, FITC-Cys-His-Ala-Ile-Tyr-Pro-Arg-His-OH) was from Kelowna International Scientific Inc. (Taipei, Taiwan). Anti-human CD71 (transferrin receptor) monoclonal antibody, allophycocyanin (APC) and mouse IgG1 kappa Isotype Control APC were from eBioscience, Inc. (Vienna, Austria). Seliciclib (purity > 99%) was from LC Laboratories (Woburn, MA, USA). A549, MDA-MB-231, SKOV-3 and U87-MG cell lines were from Bioresource Collection and Research Center (Hsinchu, Taiwan).

He G.Z, & Lin W.J. (2021). Peptide-Functionalized Nanoparticles-Encapsulated Cyclin-Dependent Kinases Inhibitor Seliciclib in Transferrin Receptor Overexpressed Cancer Cells. Nanomaterials, 11(3), 772.

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Synthesis of Iron-Phosphate-Silica Nanocomposites

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All reagents and solvents were of analytical grade and used without further purification. Iron(iii) chloride hexahydrate (FeCl₃·6H₂O, 97%), tetraethyl orthosilicate (TEOS) and ciprofloxacin (98%) were purchased from Sigma-Aldrich, while di-ammonium hydrogen phosphate ((NH₄)₂HPO₄, 99+%) was procured from Merck and ammonium hydroxide solution (NH₄OH, 28–30%) from Fluka. Cell culture materials were obtained by Life Technologies except for paraformaldehyde (PFA) (Merck) and thiazolyl blue tetrazolium bromide (MTT, 98%, Alfa Aesar).

Gessner I., Krakor E., Jurewicz A., Wulff V., Kling L., Christiansen S., Brodusch N., Gauvin R., Wortmann L., Wolke M., Plum G., Schauss A., Krautwurst J., Ruschewitz U., Ilyas S, & Mathur S. (2018). Hollow silica capsules for amphiphilic transport and sustained delivery of antibiotic and anticancer drugs. RSC Advances, 8(44), 24883-24892.

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Cell Viability and Proliferation Analysis

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Cell viability and proliferation were analyzed by Thiazolyl blue tetrazolium bromide (MTT) and Live/Dead assays. The MTT assay was performed every seven days after cell encapsulation, i.e., on days 1, 7, 14, 21, and 28. Thiazolyl blue tetrazolium bromide (Alfa Aesar, Haverhill, MA, USA) was dissolved in PBS at a concentration of 5 mg mL⁻¹ to prepare an MTT solution. The hydrogels were added to 1 mL of a fresh medium. Afterward, 0.1 mL of the MTT solution was added to the medium, followed by a 4 h incubation at 36.5 °C. After incubation, the hydrogels were placed in 1 mL of DMSO to dissolve purple formazan formed by the reduction of MTT and shaken for 30 min to mix, at which point 0.2 mL of DMSO including purple formazan was transferred to 96 wells to measure the absorbance at 570 nm.
The Live/Dead assay was conducted every seven days after cell encapsulation, i.e., on days 1, 7, and 14. A Live/Dead cell imaging kit (Invitrogen, Carlsbad, CA, USA) was used to observe the distribution of live and dead cells with a confocal laser scanning microscope (CLSM; LSM 880 with Airyscan, Carl Zeiss, Oberkochen, Germany) after fluorescent staining.

Choi J., Lee J., Shin M.E., Been S., Lee D.H, & Khang G. (2020). Eggshell Membrane/Gellan Gum Composite Hydrogels with Increased Degradability, Biocompatibility, and Anti-Swelling Properties for Effective Regeneration of Retinal Pigment Epithelium. Polymers, 12(12), 2941.

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Fluorescence Microscopy Reagent Preparation

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Dimethyl sulfoxide (DMSO), methanol (MeOH),
distilled water, and 0.05 M Tris-HCl buffer (pH 7.9) were used as
solvents. In addition, high molecular weight dsDNA (double-stranded
DNA) from Salmon testis, RNA, human serum albumin (HSA), and beta-lactoglobulin
(BLG) were purchased from Sigma-Aldrich Co. MitoTracker CMXRos Red,
LysoTracker Deep Red, Hoechst 33342, and Thiazolyl Blue tetrazolium
bromide (MTT) were purchased from ThermoFisher Scientific (Invitrogen).

Aristova D., Selin R., Heil H.S., Kosach V., Slominsky Y., Yarmoluk S., Pekhnyo V., Kovalska V., Henriques R., Mokhir A, & Chernii S. (2022). Trimethine Cyanine Dyes as NA-Sensitive Probes for Visualization of Cell Compartments in Fluorescence Microscopy. ACS Omega, 7(51), 47734-47746.

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Investigating MHY695's Effects on Cell Viability

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An MTT assay was performed to investigate the effects of MHY695 on the viabilities of NCM460, Caco-2, DLD-1, HT-29 and HCT116 cells. Thiazolyl blue tetrazolium bromide (Sigma-Aldrich; Merck KGaA) was dissolved in PBS to a stock concentration of 5 mg/ml, and further diluted with RPMI-1640 or DMEM/High glucose medium to a working concentration of 0.5 mg/ml. Cells were cultured in a 96-well cell culture plate at a density of 5×10³ cells/well for 24 h prior to treatment. The vehicle control (0.05% DMSO in culture medium) or MHY695 samples were treated with 25 or 50 µM in sextuplicate for 24 h. Thiazolyl blue tetrazolium bromide (0.5 mg/ml) was subsequently added, and the cells were incubated in the dark for 2 h. The formazan crystals were dissolved in DMSO and absorbance was measured at 540 nm using a microplate spectrophotometer (Thermo Fisher Scientific, Inc.). Each experiment was performed three times independently. The half maximal inhibitory concentration (IC₅₀) values were obtained using GraphPad Prism 5 (GraphPad Software).

Heo G., Kang D., Park C., Kim S.J., Choo J., Lee Y., Yoo J.W., Jung Y., Lee J., Kim N.D., Chung H.Y., Moon H.R, & Im E. (2019). Pro-apoptotic effect of the novel benzylidene derivative MHY695 in human colon cancer cells. Oncology Letters, 18(3), 3256-3264.

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Reagents for Cell Culture and Microscopy

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Reagents were obtained from the following manufacturers: Carl Roth GmbH & Co. KG (Karlsruhe, Germany): Agar-Agar (Kobe I), ethanol (HPLC grade), d (+)-glucose monohydrate (> 99.5%), glutaraldehyde (25%, for electron microscopy) 1,1,1,3,3,3-hexamethyldisilazane (HMDS, ≥ 98%), paraformaldehyde, sodium dodecyl sulfate (SDS, >99.5%), and sodium hydroxide (> 98%); Degussa AG (Hanau, Germany): Osmium tetroxide (75%); Sigma Aldrich Chemie GmbH (Taufkirchen, Germany): Amphotericin B, 5(6)-carboxyfluorescein (for fluorescence), dimethyl sulfoxide (DMSO, ≥ 99.9%), disodium hydrogen phosphate (> 98%), fluconazole, flucytosine, glycerol-gelatin mounting medium (GG1), poly-L-lysine solution (0.01%), sodium dihydrogen phosphate (>99%), thiazolyl blue tetrazolium bromide (MTT; ≥ 97.5%), and Triton X-100; Thermo Fisher Scientific Inc. (Darmstadt, Germany): Gibco® DMEM/F-12 medium, Gibco® PBS, Gibco® Penicillin–Streptomycin (10,000 U/mL), and Gibco® Trypsin–EDTA (0.5%); VWR International GmbH (Darmstadt, Germany): peptone (from casein).

Brakel A., Grochow T., Fritsche S., Knappe D., Krizsan A., Fietz S.A., Alber G., Hoffmann R, & Müller U. (2024). Evaluation of proline-rich antimicrobial peptides as potential lead structures for novel antimycotics against Cryptococcus neoformans. Frontiers in Microbiology, 14, 1328890.

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