INT (Iodonitrotetrazolium chloride)
(Synonyms: 碘硝基氯化四氮唑蓝,Iodonitrotetrazolium chloride; p-Iodonitrotetrazolium Violet) 目录号 : GC30118
A monotetrazolium indicator dye
Cas No.:146-68-9
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Iodonitrotetrazolium (INT) is a monotetrazolium salt used as an indicator dye.1,2 It is reduced to an insoluble formazan that is used as a vital dye or indicator of cellular redox activity.1 Reduction commonly results from the activity of dehydrogenases, although non-enzymatic electron transfer reactions can occur in the presence of an intermediate electron acceptor.1,3 INT is commonly used to measure the respiratory activity of microorganisms in a variety of contexts.1,2
1.Berridge, M.V., Tan, A.S., and Herst, P.M.Tetrazolium dyes as tools in cell biology: New insights into their cellular reductionBiotechnol. Ann. Rev.11127-152(2005) 2.Sabnis, R.W.Handbook of biological dyes and stains: Synthesis and industrial applications(2010) 3.Green, J.D., and Narahara, H.T.Assay of succinate dehydrogenase activity by the tetrazolium method: Evaluation of an improved technique in skeletal muscle fractionsJ. Histochem. Cytochem.28(5)408-412(1980)
| Cas No. | 146-68-9 | SDF | |
| 别名 | 碘硝基氯化四氮唑蓝,Iodonitrotetrazolium chloride; p-Iodonitrotetrazolium Violet | ||
| Canonical SMILES | IC1=CC=C(N2N=C(C3=CC=CC=C3)N=[N+]2C4=CC=C([N+]([O-])=O)C=C4)C=C1.[Cl-] | ||
| 分子式 | C19H13ClIN5O2 | 分子量 | 505.7 |
| 溶解度 | DMSO : ≥ 42 mg/mL (83.05 mM) | 储存条件 | Store at -20°C |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 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。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 1.9775 mL | 9.8873 mL | 19.7746 mL |
| 5 mM | 0.3955 mL | 1.9775 mL | 3.9549 mL |
| 10 mM | 0.1977 mL | 0.9887 mL | 1.9775 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
Comparison of the fluorescent redox dye 5-cyano-2,3-ditolyltetrazolium chloride with p-iodonitrotetrazolium violet to detect metabolic activity in heat-stressed Listeria monocytogenes cells
J Appl Bacteriol 1994 Oct;77(4):353-8.7989262 10.1111/j.1365-2672.1994.tb03434.x
The fluorogenic redox indicator 5-cyano-2,3-ditolyltetrazolium chloride (CTC) was compared with the chromogenic p-iodonitrotetrazolium violet (INT) and conventional methods to assess cellular viability. Mild heat treatment was used as well-controlled method for producing non-viable and sub-lethally injured cells. CTC gave an underestimation of the viability of Listeria monocytogenes cells when compared with classical plating methods whereas INT gave an overestimation. However, CTC proved to be a sensitive indicator of uninjured cells. The difference between the total count and the CTC count was equivalent to the injured cell population. The fluorescent formazan formed on reduction of CTC was readily detected with a charge coupled device and cells enumerated automatically using image analysis.
Antibacterial secondary metabolites from Vernonia auriculifera Hiern (Asteraceae) against MDR phenotypes
Nat Prod Res 2022 Jun;36(12):3203-3206.34293972 10.1080/14786419.2021.1953024
Purification of the aerial parts of Vernonia auriculifera Hiern afforded steroids (1-2), flavonoids (3-5), and polyalcohol (6). Their structures were determined using spectral evidences as well as by comparison with reported data. Iodonitrotetrazolium chloride (INT) colorimetric assay was used to assess the antibacterial activity of the extract and isolates against 13 pathogenic strains. The crude extract showed strong antibacterial activity (MIC < 100 µg/mL) against the tested bacterial strains. When combined with an efflux pump inhibitor phenylalanine beta naphthylamide (PAβN), the inhibition potency of the extract was substantially enhanced with the lowest MIC value at 4 µg/mL. Compounds 5 and 6 showed moderate activity (MIC < 100 µg/mL) against 12/13 (92.3%), and 8/13 (61.5%) bacterial strains, respectively. A minimal bactericidal concentration (MBC)/minimal inhibitory concentration (MIC) ratio ≿4 indicated their bactericidal effect against Escherichia coli, Enterbacter aerogenes, Klebsiella pneumoniae, Providencia stuartii, Pseudomonas aeruginosa, and Staphylococcus aureus.
INT (2-(4-Iodophenyl)-3-(4-Nitrophenyl)-5-(Phenyl) Tetrazolium Chloride) Is Toxic to Prokaryote Cells Precluding Its Use with Whole Cells as a Proxy for In Vivo Respiration
Microb Ecol 2015 Nov;70(4):1004-11.25991603 10.1007/s00248-015-0626-3
Prokaryote respiration is expected to be responsible for more than half of the community respiration in the ocean, but the lack of a practical method to measure the rate of prokaryote respiration in the open ocean resulted in very few published data leaving the role of organotrophic prokaryotes open to debate. Oxygen consumption rates of oceanic prokaryotes measured with current methods may be biased due to pre-incubation size filtration and long incubation times both of which can change the physiological and taxonomic profile of the sample during the incubation period. In vivo INT reduction has been used in terrestrial samples to estimate respiration rates, and recently, the method was introduced and applied in aquatic ecology. We measured oxygen consumption rates and in vivo INT reduction to formazan in cultures of marine bacterioplankton communities, Vibrio harveyi and the eukaryote Isochrysis galbana. For prokaryotes, we observed a decrease in oxygen consumption rates with increasing INT concentrations between 0.05 and 1 mM. Time series after 0.5 mM INT addition to prokaryote samples showed a burst of in vivo INT reduction to formazan and a rapid decline of oxygen consumption rates to zero within less than an hour. Our data for non-axenic eukaryote cultures suggest poisoning of the eukaryote. Prokaryotes are clearly poisoned by INT on time scales of less than 1 h, invalidating the interpretation of in vivo INT reduction to formazan as a proxy for oxygen consumption rates.
Effects of substrates and phosphate on INT (2-(4-iodophenyl)-3-(4-nitrophenyl)-5-phenyl tetrazolium chloride) and CTC (5-cyano-2,3-ditolyl tetrazolium chloride) reduction in Escherichia coli
J Appl Bacteriol 1996 Feb;80(2):209-15.8642015 10.1111/j.1365-2672.1996.tb03212.x
The effects of substrates of primary aerobic dehydrogenases, and inorganic phosphate on aerobic INT and CTC reduction in Escherichia coli were examined. In general, INT produced less formazan than CTC, but INT (+) cell counts remained near values of CTC (+) cells. INT and CTC (+) cell numbers were higher than plate counts on R2A medium using succinate, formate, lactate, casamino acids, glucose, glycerol (INT only) and no substrate. Formate resulted in the greatest amount of INT and CTC formazan. Reduction of both INT and CTC was inhibited above 10 mmol l-1 phosphate, and this appeared to be related to decreased rates of O2 consumption. Formation of fluorescent CTC (+), but not INT (+) cells was also inhibited in a concentration dependent manner by phosphate above 10 mmol l-1. From light microscopic observations it appeared CTC formed increasing amounts of poorly or non-fluorescent formazan with increasing phosphate. Therefore, use of phosphate buffer in excess of 10 mmol l-1 may not be appropriate in CTC and INT reduction assays.
Bioprospecting of endophytic actinobacterium associated with Aloe ferox mill for antibacterial activity
BMC Complement Med Ther 2022 Oct 3;22(1):258.36192707 PMC9531469
Background: The emergence of drug resistance among pathogens has resulted in renewed interest in bioprospecting for natural microbial products. Methods: This study aimed to bioprospecting endophytic actinobacterium associated with Aloe ferox Mill for its antibacterial activity. Endophytic actinomycetes were isolated from the gel of A. ferox Mill by surface sterilization technique using actinomycete isolation agar. The isolate with a promising antibacterial activity was identified using 16S rRNA sequence analysis. The minimum inhibitory concentration (MIC) of the extract was assessed by the micro-dilution method and its effect on the respiratory chain dehydrogenase (RCD) activity was ascertained by the Iodonitrotetrazolium chloride (INT) assay. Fourier transform-infrared spectrophotometer (FTIR) and gas chromatography-mass spectrophotometry (GC-MS) were employed to identify functional groups and the chemical constituents, respectively. Results: The actinobacterium was found to be Streptomyces olivaceus CP016795.1. Its extract displayed noteworthy antibacterial activity (MIC ≿ mg/mL) against Staphylococcus aureus (ATCC 25925), Bacillus cereus (ATCC 10102), and Escherichia coli (ATCC 25922); and showed an inhibitory effect on the RCD activity. FTIR spectrum displayed hydroxyl, amine, and aromatic groups, and the GC-MS revealed 5-Hydroxymethylfurfural as the main constituent (19.47%). Conclusions: S. olivaceus CP016795.1 can serve as a potential source of effective antibacterial compounds.