Mono-sulfonated tetrazolium salt based NAD(P)H detection reagents suitable for dehydrogenase and real-time cell viability assays

Zhang, Wei; Zhu, Min; Wang, Feng; Cao, Danhui; Ruan, Jennifer Jin; Su, Weike; Ruan, Benfang

doi:10.1016/j.ab.2016.06.026

Cited by 27 publications

(31 citation statements)

References 21 publications

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“…Because of its simple protocol, the WST-8 (CCK-8) assay has become a popular cell vitality method [78][79][80]. However, WST-8 is not very stable especially under the reduced condition, so EZMTT was developed for drug-induced proliferation assay [81]. EZMTT is another water-soluble tetrazolium salt which is less toxic than also, EZMTT reagent showed excellent stability and signal of background ratio.…”

Section: Nadh and Nadph Production Of Live Cellmentioning

confidence: 99%

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Cell Growth Measurement

Chen¹,

Rui²,

Yu³

et al. 2020

Cell Growth

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The cell is the basic structural and functional unit of all living organisms. As the smallest unit and building blocks of life, cells differ in size, shape, metabolism, reproduction, and growth requirements. Cells reproduce through cell division involving a four-phase (G1, S, G2, M) cell cycle, which is tightly regulated at multiple checkpoints. The resulting growth curve demonstrates that cell population increases in three sequential steps: incubation, exponential hyperplasia, and stagnation/death phases. Cell growth is subject to changes in disease state and/or environmental conditions. This chapter will focus on methods for cell growth measurement, which are grouped into five sections: cell cycle, apoptosis, growth curve, druginduced proliferation (DIP), and continuous assays. Among the continuous assays, the EZMTT dye allows for long-term tracking of cell growth under various conditions and shows promise in precision medicine by early detection of drug resistance.

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Section: Nadh and Nadph Production Of Live Cellmentioning

confidence: 99%

“…The EZMTT dye [81] was initially designed to overcome the stability issue of WST-8 (CCK-8) reagent which can cause false positives in the presence of antioxidants such as BME (Figure 5A) or EGCG. Later, the EZMTT dye was found to be essentially nontoxic and stable in various media [102].…”

Section: Ezmtt Dye-based Cell Proliferation Analysismentioning

confidence: 99%

Cell Growth Measurement

Chen¹,

Rui²,

Yu³

et al. 2020

Cell Growth

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“…Therefore, it is critically important to develop a sensitive continuous assay that reliably detects minor growth after drug treatment. Since the EZMTT reagent was shown to have high sensitivity and is capable of tracking time-dependent cell growth 12,13 , we further developed the EZMTT-based continuous assays to measure the DIP rate for infectious bacteria. As expected, the assay provided rapid, reproducible and precise assessment of drug-resistance and importantly, reliable detected10-30% more partial drug resistant bacteria than the current turbidity-based diagnostic method..…”

Section: Introductionmentioning

confidence: 99%

Detection of “Hidden” Antimicrobial Drug Resistance

Hu¹,

et al. 2019

ACS Infect. Dis.

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“…ZoIs of bacterial growth are visualised using metabolic stains, typically tetrazolium salts such as 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT) or 2‐(4‐iodophenyl)‐3‐(4‐nitrophenyl)‐5‐phenyl‐2 H ‐tetrazolium chloride (INT) . These salts are reduced by dehydrogenases in viable cells to produce purple formazan, thus ZoIs are visualised as white spots on a purple background . Lead compounds are then extracted and characterised.…”

Section: Introductionmentioning

confidence: 99%

“…[20][21][22][23][24] These salts are reduced by dehydrogenases in viable cells to produce purple formazan, thus ZoIs are visualised as white spots on a purple background. 24,25 Lead compounds are then extracted and characterised.…”

Section: Introductionmentioning

confidence: 99%

Characterisation and screening of antimicrobial essential oil components against clinically important antibiotic‐resistant bacteria using thin layer chromatography‐direct bioautography hyphenated with GC‐MS, LC‐MS and NMR

Owen

White

Laird

2018

Phytochemical Analysis

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Introduction The antimicrobial activity of many essential oils (EOs) is well established, indicating that EOs may be a source of compounds for antimicrobial drug development. Thin layer chromatography‐direct bioautography (TLC‐DB) can quickly identify antimicrobial components in complex mixtures and can be applied to the screening of EOs for lead compounds. Objectives This study aimed to identify antimicrobial components of oregano, rosewood and cumin EOs against antibiotic‐sensitive and ‐resistant bacteria using TLC‐DB and a multi‐faceted approach of GC‐MS, LC‐MS and NMR techniques to characterise bioactive compounds. The study also aimed to quantify the antimicrobial activity of bioactive compounds in order to evaluate their potential for the development of therapies against antibiotic‐resistant bacteria. Materials and Methods EOs were eluted on TLC plates and sprayed with a suspension of Staphylococcus aureus, Enterococcus faecium, Escherichia coli or Pseudomonas aeruginosa (antibiotic‐sensitive and ‐resistant isolates). Zones of inhibition, visualised with iodonitrotetrazolium chloride, were subject to GC‐MS, LC‐MS and NMR to characterise the bioactive compounds. Results Seven compounds were identified from the three EOs using GC‐MS, while LC‐MS and NMR failed to detect the presence of any further non‐volatile or heat labile compounds. Carvacrol was most antimicrobial compound identified, with minimum inhibitory concentrations ranging 0.99–31.62 mM. Conclusion The identified antimicrobial compounds present in oregano, rosewood and cumin EOs including carvacrol may be candidates for the development of novel antimicrobial therapies against antibiotic‐resistant bacteria.

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Mono-sulfonated tetrazolium salt based NAD(P)H detection reagents suitable for dehydrogenase and real-time cell viability assays

Cited by 27 publications

References 21 publications

Cell Growth Measurement

Cell Growth Measurement

Detection of “Hidden” Antimicrobial Drug Resistance

Characterisation and screening of antimicrobial essential oil components against clinically important antibiotic‐resistant bacteria using thin layer chromatography‐direct bioautography hyphenated with GC‐MS, LC‐MS and NMR

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