• Title/Summary/Keyword: Colorimetric assay

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The Growth Inhibitory Effects of Epigallocatechin Gallate Against Human Skin Melanoma Cells and Human Oral Epitheloid Carcinoma Cells (Epigallocatechin gallate의 인체 피부흑색종세포와 인체 구강유상피암종세포에 대한 성장억제효과)

  • 한두석;박승택;백승화
    • Environmental Mutagens and Carcinogens
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    • v.18 no.2
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    • pp.98-103
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    • 1998
  • Epigallocatechin gallate (EGCG) was reported to exert weak cytotoxicity against normal healthy cells such as C3H10T1/2 cells, but profound inhibitory effects on the initiation or promotion stage of chemical carcinogenesis in mammary gland, blood and mouse skin. This study was carried out to develop antitumor agents with weak side effects and strong antitumor activity. Human skin melanoma cells (HBT 69) and human oral epitheloid carcinoma cells (OCL 17) were cultured in RPMI-1640 media containing 10% fetal bovine serum, antibiotic, and fungizone. After incubation for 24 hrs, the cells were treated with various amounts of (EGCG) for 48 hrs. The growth inhibitory effects of EGCG in human oral epitheloid carcinoma cells were evaluated by the 3- (4,5-djmethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), neutral red (NR), and sulforhodamine B protein (SRB) assays of colorimetric methods. The light microscopic study was also carried out to observe morphological changes of the treated cells. These results obtained were as follows; 1. Significantly inhibitory effects of EGCG against cultured human oral epithelioid carcinoma cells. 2. Significantly inhibitory effects against cultured human skin melanoma cells treated with 50 $\mu$M EGCG, but decreased inhibitory effects in 100 $\mu$M EGCG. 3. Degenerative changes against cultured human oral epitheloid carcinoma cells. 4. Degenerative changes against human skin melanoma cells treated with 50 UM EGCG, but recovered degenerative changes in 100 $\mu$M EGCG.

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Colorimetric Evaluation of the Time-Killing Assay for Citropin 1.1, Lipopeptide Palm-KK-$NH_2$, and Temporin A

  • Baranska-Rybak, Wioletta;Dawgul, Malgorzata;Bielinska, Sylwia;Kraska, Bartlomiej;Piechowicz, Lidia;Kamysz, Wojciech
    • Journal of Microbiology and Biotechnology
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    • v.21 no.5
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    • pp.536-539
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    • 2011
  • Nowadays, there are a number of colorimetric techniques available for the determination of a time killing assay in a manner much easier and faster than those previously more commonly used, which were much more time-consuming and laborious colony counting procedures. Here, an attempt has been made to test the antimicrobial peptides of Citropin 1.1, Palm-KK-$NH_2$, and Temporin A on a reference strain of Staphylococcus aureus using resazurin as the cell viability reagent. Staphylococcus aureus was exposed to the test compounds over varying periods of time and the metabolic activity measured, with a profile of antimicrobial activity then established. The results are in agreement with data from previous literature, thus confirming the relevance of the application of resazurin for the testing of antimicrobial agents.

Enzyme-linked Immunosorbent Assay Strip Sensor for Rapid Detection of Staphylococcus aureus (Staphylococcus aureus 신속 검출을 위한 효소면역측정 스트립 센서)

  • Park, So Jung;Kim, Young-Kee
    • Applied Chemistry for Engineering
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    • v.22 no.5
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    • pp.522-525
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    • 2011
  • In this study, an established enzyme-linked immunosorbent assay and immuno-chromatography technique are combined to fabricate an immuno-strip sensor for the detection of S. aureus. The immuno-strip is manufactured by using four different functional membranes. The capture antibody is immobilized on the nitrocellulose membrane due to the high affinity and the capillary action through porous membranes induces a flow of sample. A colorimetric signal is appeared according to the enzyme reaction and is analyzed by the digital camera (qualitative analysis) and home-made image analysis software (quantitative analysis). Under the optimal conditions, samples with S. aureus in the range of $2.7{\times}10^4{\sim}2.7{\times}10^7CFU/mL$ can be detected by the colorimetric method within 30 min.

Phenolic Compounds from Duchesnea chryszntha and their Cytotoxic Activities in Human Cancer Cell

  • Lee, Ihn-Rhan;Yang, Mi-Young
    • Archives of Pharmacal Research
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    • v.17 no.6
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    • pp.476-479
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    • 1994
  • Five pohenolic compounds were isolated from 80% aq. acetone extract of Duchesnea chyrysantha. Their crytotoxicities were screened by the colorimetric tetrazolium assay (MIT assay). Gallic acid, methyl caffeate, protocatechuic acid and pedunculagin mildly inhibited the survival of $PC_{14}{\;}and{\;}MKN_{45}$ human cancer cell. Brevifolin carboxylic acid showed a strong cytotoxic activity.

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The Inhibitory Effects of Taraxaci Herba against Cadmium induced Cytotoxicity (포공령의 카드뮴에 대한 세포독성 억제효과)

  • Han, Du-Seok;Lee, Ki-Nam;Lee, Jong-Sub;Baek, Seung-Hwa
    • YAKHAK HOEJI
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    • v.42 no.3
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    • pp.307-311
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    • 1998
  • This study was carried out to evaluate antitoxic effects Taraxaci Herba extract against Cadium by calorimetric methods. The antitoxic activity of Taraxaci Herba ex tract in NIH 3T3 fibroblasts was evaluated by MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-phenyl-2H-tetrazoliumbromide), NR (Neutral red) and SRB (Sulforhodamine B protein) assay. The light microscopic study was carried out to observe morphological changes of the treated cells. These results were obtained as follows; The concentration of $10^{-2}mg/ml$ of Taraxaci Herba extract was shown significant antitoxic activity. The number of NIH 3T3 fibroblasts were antitoxic and tend to regenerate. These results suggest that Taraxaci Herba extract retains a potential antitoxic activity.

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A STUDY ON THE CYTOTOXIC EFFECTS OF MITOMYCIN C AND 5-FLUOROURACIL IN CULTURED RAT FIBROBLASTS

  • C. S. M;Park, Hong-Seog;Chung, Yeun-Tai
    • Toxicological Research
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    • v.7 no.1
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    • pp.13-20
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    • 1991
  • To investigate the cytotoxicity and genotoxicity of the DNA alkylating agnet, mitomycin C and the antimetabolite, 5-Fluorouracil (5-FU) in cultured rat fibroblasts, the colorimetric assay of netural red (NR) for cytotoxicity and for genotoxicity, sister chromatid exchange (SCE) assay and the measurement of the rate of DNA synthesis were performed in cells cultured in media containing various concentrations of mitomycin C and 5-FU. The uptake ability of neutral red decreased does-dependently. NR90 and NR50 values of mitomycin C were 1.49 nM and 6.87mM and 5-FU were 38.4mM AND 284.4Mm respectively.

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Salicylimine-Based Colorimetric and Fluorescent Chemosensor for Selective Detection of Cyanide in Aqueous Buffer

  • Noh, Jin Young;Hwang, In Hong;Kim, Hyun;Song, Eun Joo;Kim, Kyung Beom;Kim, Cheal
    • Bulletin of the Korean Chemical Society
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    • v.34 no.7
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    • pp.1985-1989
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    • 2013
  • A simple colorimetric and fluorescent anion sensor 1 based on salicylimine showed a high selectivity and sensitivity for detection of cyanide in aqueous solution. The receptor 1 showed high selectivity toward $CN^-$ ions in a 1:1 stoichiometric manner, which induces a fast color change from colorless to orange and a dramatic enhancement in fluorescence intensity selectively for cyanide anions over other anions. Such selectivity resulted from the nucleophilic addition of $CN^-$ to the carbon atom of an electron-deficient imine group. The sensitivity of the fluorescence-based assay (0.06 ${\mu}M$) is below the 1.9 ${\mu}M$ suggested by the World Health Organization (WHO) as the maximum allowable cyanide concentration in drinking water, capable of being a practical system for the monitoring of $CN^-$ concentrations in aqueous samples.

Development of Colorimetric Paper Sensor for Pesticide Detection Using Competitive-inhibiting Reaction

  • Kim, Hyeok Jung;Kim, Yeji;Park, Su Jung;Kwon, Chanho;Noh, Hyeran
    • BioChip Journal
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    • v.12 no.4
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    • pp.326-331
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    • 2018
  • Contamination by pesticides is an everincreasing problem associated with fields of environmental management and healthcare. Accordingly, appropriate treatments are in demand. Pesticide detection methods have been researched extensively, aimed at making the detection convenient, fast, cost-effective, and easy to use. Among the various detecting strategies, paper-based assay is potent for real-time pesticide sensing due to its unique advantages including disposability, light weight, and low cost. In this study, a paper-based sensor for chlorpyrifos, an organophosphate pesticide, has been developed by layering three sheets of patterned plates. In colorimetric quantification of pesticides, the blue color produced by the interaction between acetylcholinesterase and indoxyl acetate is inhibited by the pesticide molecules present in the sample solutions. With the optimized paper-based sensor, the pesticide is sensitively detected (limit of detection =8.60 ppm) within 5min. Furthermore, the shelf life of the device is enhanced to 14 days after from the fabrication, by treating trehalose solution onto the deposited reagents. We expect the paper-based device to be utilized as a first-screening analytic device for water quality monitoring and food analysis.

A Neutravidin-based Assay for Reverse Transcriptase Suitable for High Throughput Screening of Retroviral Activity

  • Brennan, Lyndall E.;Sune, Carlos;Klimkait, Thomas
    • BMB Reports
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    • v.35 no.3
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    • pp.262-266
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    • 2002
  • A non-isotopic neutravidin-based reverse transcriptase (RT) assay adapted for high throughput screening of HIV activity is described. Using a 96-well microtitre plate, HIV particles are lysed and the RT enzyme released into a reaction mixture containing poly(A) RNA, biotinylated oligo d(T) and fluorescein-labelled dUTP (FI-dUTP). With poly(A) as a template and oligo d(T) as primer, the viron RT incorporates FI-dUTP into an elongating DNA strand. The resulting product is captured on a neutravidin-coated 96-well plate and the unincorporated nucleotides removed by a series of washing steps. A simple ELISA is subsequently performed using a monoclonal antifluorescein antibody conjugated to alkaline phosphatase. Quantification of RT activity is facilitated by a colorimetric readout. The assay was validated in the context of a diagnostic HIV-1 phenotyping assay. Using supernatants from HIV-1 infected lymphocyte cultures the assay was shown to be as sensitive as a radioactive assay and the RT activity correlated well with levels of cell-asociated HIV-p24. Importantly, even minor reductions of RT activity by virus variants with reduced fitness could be distinguished.