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5-Cyano-2,3-di-(p-tolyl)tetrazolium (chloride)

(Synonyms: 5-氰基-2,3-二(4-甲基苯基)四唑氯化物,CTC) 目录号 : GC42502

A redox-sensitive fluorescent probe

5-Cyano-2,3-di-(p-tolyl)tetrazolium (chloride) Chemical Structure

Cas No.:90217-02-0

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产品描述

5-Cyano-2,3-di-(p-tolyl)tetrazolium (chloride) is a redox-sensitive tetrazolium salt used primarily to detect metabolic activity in microorganisms. This membrane-permeable indicator is readily taken up by living cells and reduced by respiratory activity to produce a red fluorescent, water-insoluble formazan crystal (absorption maximum at 450 nm). While the oxidized salt is non-fluorescent, the reduced formazan can be evaluated using a standard rhodamine filter set (excitation: 510-560 nm; emission > 590 nm), although optimum excitation requires a blue 420 nm excitation filter. For flow cytometry, a 480 nm excitation laser combined with detection in the red region is suitable.

Chemical Properties

Cas No. 90217-02-0 SDF
别名 5-氰基-2,3-二(4-甲基苯基)四唑氯化物,CTC
Canonical SMILES CC(C=C1)=CC=C1N2N=C(C#N)N=[N+]2C3=CC=C(C)C=C3.[Cl-]
分子式 C16H14N5•Cl 分子量 311.8
溶解度 DMF: 0.1 mg/ml,DMSO: 0.25 mg/ml,Ethanol: 3 mg/ml,PBS (pH 7.2): 1 mg/ml 储存条件 Store at -20°C; protect from light
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储备液的保存方式和期限:-80°C 储存时,请在 6 个月内使用,-20°C 储存时,请在 1 个月内使用。
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1 mM 3.2072 mL 16.0359 mL 32.0718 mL
5 mM 0.6414 mL 3.2072 mL 6.4144 mL
10 mM 0.3207 mL 1.6036 mL 3.2072 mL
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Research Update

Tetrazolium salts and formazan products in Cell Biology: Viability assessment, fluorescence imaging, and labeling perspectives

Acta Histochem 2018 Apr;120(3):159-167.PMID:29496266DOI:10.1016/j.acthis.2018.02.005.

For many years various tetrazolium salts and their formazan products have been employed in histochemistry and for assessing cell viability. For the latter application, the most widely used are 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), and 5-cyano-2,3-di-(p-tolyl)-tetrazolium chloride (CTC) for viability assays of eukaryotic cells and bacteria, respectively. In these cases, the nicotinamide-adenine-dinucleotide (NAD(P)H) coenzyme and dehydrogenases from metabolically active cells reduce tetrazolium salts to strongly colored and lipophilic formazan products, which are then quantified by absorbance (MTT) or fluorescence (CTC). More recently, certain sulfonated tetrazolium, which give rise to water-soluble formazans, have also proved useful for cytotoxicity assays. We describe several aspects of the application of tetrazolium salts and formazans in biomedical cell biology research, mainly regarding formazan-based colorimetric assays, cellular reduction of MTT, and localization and fluorescence of the MTT formazan in lipidic cell structures. In addition, some pharmacological and labeling perspectives of these compounds are also described.

Differential resuscitative effect of pyruvate and its analogues on VBNC (viable but non-culturable) Salmonella

Microbes Environ 2013;28(2):180-6.PMID:23595023DOI:10.1264/jsme2.me12174.

An environmental isolate of Salmonella Enteritidis (SE), grown to the logarithmic phase, rapidly lost culturability by the addition of 3 mM H2O2 to cultures grown in Luria-Bertani (LB) medium; however, some H2O2-treated bacteria regained their culturability in M9 minimal medium, if sodium pyruvate was present at at least 0.3 mM. In addition, most pyruvate analogues, such as bromopyruvate or phenylpyruvate, did not show restoration activity similar to that of pyruvate, except in the case of α-ketobutyrate. Further analysis of the mechanism underlying the resuscitation by pyruvate revealed that although many of the bacteria showed respiratory activity on CTC (5-cyano-2,3-di-(p-tolyl) tetrazolium chloride) reduction with or without pyruvate, the biosynthesis of DNA and protein synthesis were quite different in the presence or absence of pyruvate, i.e., pyruvate endowed the cells with the ability to incorporate much more radio-label into precursors during the resuscitation process. These results suggest that pyruvate is one of the key molecules working in the resuscitation process by taking bacteria from the non-culturable state to the growing and colony-forming state by triggering the synthesis of macromolecules such as DNA and protein.

Induction of viable but non-culturable (VBNC) state in Salmonella cultured in M9 minimal medium containing high glucose

Biol Pharm Bull 2014;37(10):1617-25.PMID:25109306DOI:10.1248/bpb.b14-00322.

An environmental isolate of Salmonella enterica serovar Enteritidis (SE) clone, SE Cl#15-1, loses its culturability during 72-h culture in M9 minimal medium containing 0.8% glucose, a concentration twice higher than that in normal M9 medium, whereas the bacterium retains its culturability in normal M9 medium. Live/dead analysis using the 5-cyano-2,3-di(p-tolyl) tetrazolium chloride (CTC)-reduction assay revealed that SE cells cultured in M9 medium containing 0.8% glucose died with time when in the "viable but non-culturable" (VBNC) state. Assay of the culturability of SE cells in the used supernatant (0.4 spent M9 or 0.8 spent M9) also indicated that 0.8 spent M9 soon showed a lethal effect on intact SE cells. These results suggest that large amounts of glucose metabolites might have been responsible for the toxicity. Analysis of the 0.8 spent M9 revealed that formate rapidly accumulated in the medium. The pH of the medium rapidly dropped to 4.7, leading to conversion of formate to formic acid, which might have damaged the bacterial cell membrane. These results suggest that the excessive amount of glucose in the M9 medium might have injured SE cells in the VBNC state by being metabolized to formic acid and other acidic compounds.