Methylthiazol tetrazolium

Manufactured by Merck Group

Sourced in United States

Methylthiazol tetrazolium is a colorimetric assay used to measure cell metabolic activity. It is a widely used lab equipment for various applications in cell biology and biochemistry research.

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Close spelling variants of this product

3,4,5-dimethyl-N-methylthiazol-2-yl-2,5-d-phenyl tetrazolium bromide (MTT) (2 citations) Tetrazolium dye 3-(4,5-di-methylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), (2 citations) Methylthiazol-2-yl-2,5-diphenyl tetrazolium bromide (MTT), (2 citations)

6 protocols using methylthiazol tetrazolium

TLC-Bioautography for Antimicrobial Screening

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One milliliter of representative MDR bacterial strains with 1.5 × 10⁸ CFU/mL concentration was used for every 10 mL of Mueller-Hinton agar. Developed TLC plates were placed in a sterile Petri dish (150 mm). The culture was added to the melted Mueller-Hinton agar and a thin layer was poured over the TLC plate. After the solidification of the medium, TLC plate was incubated for 24 hours at 35 ± 2°C. The TLC-bioautography plates were sprayed with an aqueous solution (2.5 mg/mL) of methylthiazol tetrazolium (Sigma, USA). Clear zone of inhibition was observed against a purple background [14 (link)]. Identification of specific compounds was limited by the unavailability of prepared reference standards.

Valle DL J.r., Puzon J.J., Cabrera E.C, & Rivera W.L. (2016). Thin Layer Chromatography-Bioautography and Gas Chromatography-Mass Spectrometry of Antimicrobial Leaf Extracts from Philippine Piper betle L. against Multidrug-Resistant Bacteria. Evidence-based Complementary and Alternative Medicine : eCAM, 2016, 4976791.

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Evaluating Anchorage-Independent Growth and Colony Formation of RET Mutants

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To examine anchorage-independent growth, NIH3T3 cells (2 × 10⁵ cells per well) transfected with EV, WT or RET mutants were suspended in 0.35% agar (#214220, BD Difco) containing RPMI 1640 medium with 10% fetal bovine serum and penicillin/streptomycin and seeded on 0.7% agar with the same complete media in 6-well plates. Cells were allowed to grow for 5 weeks and stained with methyl thiazol tetrazolium (#M5655, Sigma-Aldrich) for 3 h. The number of colonies was counted manually. This assay was performed in triplicate.
For plate clone formation assay, A2780 cells (500 cells per well) transfected with EV, WT or RET mutants were seeded in 6-well plates in triplicate for 14 days. Cells were fixed with methanol for 15 min and stained with crystal violet for 30 min. The number of colonies was counted manually. This experiment was performed three times.

Guan L., Li Z., Xie F., Pang Y., Zhang C., Tang H., Zhang H., Chen C., Zhan Y., Zhao T., Jiang H., Jia X., Wang Y, & Lu Y. (2020). Oncogenic and drug-sensitive RET mutations in human epithelial ovarian cancer. Journal of Experimental & Clinical Cancer Research : CR, 39, 53.

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Synthesis and Characterization of MPEG-Succinic Acid Conjugates

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Methoxypoly(ethylene glycol) (MPEG, M_n=2,000, Aldrich), succinic anhydride (SA, 99%, Alfa Aesar), 3-amino-1-propanol (AP, 99%, Alfa Aesar), hexane-1,6-dioldiacrylate (HDD, 99%), methylthiazoltetrazolium (MTT), and phosphate-buffered saline (PBS) were purchased from Sigma Chemical Co., and 1st BASE (Malaysia), respectively. N,N′-dicyclohexylcarbodiimide (DCC, 99%, Alfa Aesar), 4-(dimethyl amino)pyridine (DMAP, 99%, Alfa Aesar), cholesteryl chloroformate (Chol, 99%, Alfa Aesar), anhydrous dichloromethane (DCM), anhydrous triethylamine (TEA), 1,4-dioxane, and other materials were received and prepared as described in previous work.31 (link)

Zhang C.Y., Xiong D., Sun Y., Zhao B., Lin W.J, & Zhang L.J. (2014). Self-assembled micelles based on pH-sensitive PAE-g-MPEG-cholesterol block copolymer for anticancer drug delivery. International Journal of Nanomedicine, 9, 4923-4933.

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Doxorubicin Delivery via Albumin Nanoparticles

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Doxorubicin hydrochloride (DOX-HCl) was purchased from Wuhan Yuan Cheng Gong Chuang Co. Bovine serum albumin (BSA, ≥98%), glutaric acid (≥99%), glutaraldehyde, triethylamine (TEA, ≥99%) and methylthiazoltetrazolium (MTT) were purchased from Sigma (St. Louis, MO). Dulbecco’s modified eagle media (DMEM) growth media, fetal bovine serum (FBS), trypsin, penicillin and streptomycin, were all purchased from Invitrogen; NIH 3T3, A549, A2780 and NCL-H460 cell lines were obtained from the American Type Culture Collection (ATCC). All other reagents were used as received.

Yang Z., Zhang N., Ma T., Liu L., Zhao L, & Xie H. (2020). Engineered bovine serum albumin-based nanoparticles with pH-sensitivity for doxorubicin delivery and controlled release. Drug Delivery, 27(1), 1156-1164.

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Synthesis and Characterization of Polymeric Nanoparticles

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2-Ethyl-2-oxazoline (EOz), SPM, fluorescamine and methylthiazol tetrazolium (MTT) were purchased from Sigma (St. Louis, MO, USA). d,l-Lactic acid (d,l-LA) was from Daigang Biological Engineering Co., Ltd (Jinan, China). N-Hydroxy succinimide (NHS) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC·HCl) were from Qiyun Biotechnology Co., Ltd (Guangzhou, China). PTX (>98%) was obtained from Ruiyong Biotechnology Co., Ltd (Shanghai, China). Roswell Park Memorial Institute medium 1640 (RPMI 1640) and fetal bovine serum (FBS) were purchased from Gibco BRL Life Technologies (Gaithersburg, MD, USA). Sodium azide (NaN₃), nystatin and α-difluoromethyl ornithine (DFMO) were from Xinding pengfei Technology Development Co., Ltd. (Beijing, China), Jianglai Biological Technology Co., Ltd. (Shanghai, China) and MedChemExpress Co., Ltd. (New Jersey, USA), respectively. Lyso-Tracker Red was provided by Molecular Probes (Life Technologies, Thermo Fisher Scientific, USA).

Chen Y., Yang C., Mao J., Li H., Ding J, & Zhou W. (2019). Spermine modified polymeric micelles with pH-sensitive drug release for targeted and enhanced antitumor therapy. RSC Advances, 9(20), 11026-11037.

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Comparative Morphology and Growth Kinetics

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Both cell types, DOK^WT and early neoplastic oral keratinocyte luciferase‐transduced (DOK^Luc), were cultured at passages (45–48) and their morphology was compared under a light microscope (Nikon TS100; Nikon, Tokyo, Japan). The growth rate was analyzed using a colorimetric assay based on methylthiazol tetrazolium (Sigma) and measured at 570 nm using a microplate reader (BMG LABTECH, GmbH, Ortenberg, Germany).

Suliman S., Parajuli H., Sun Y., Johannessen A.C., Finne–Wistrand A., McCormack E., Mustafa K, & Costea D.E. (2015). Establishment of a bioluminescence model for microenvironmentally induced oral carcinogenesis with implications for screening bioengineered scaffolds. Head & Neck, 38(Suppl 1), E1177-E1187.

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