• Title/Summary/Keyword: Microplate

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Determination of Carbon Source Utilization of Bacillus and Pythium Species by Biolog$^{(R)}$ Microplate Assay

  • Chun, Se-Chul;R.W. Schneider;Chung, Ill-Min
    • Journal of Microbiology
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    • v.41 no.3
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    • pp.252-258
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    • 2003
  • The carbon utilizations of Bacillus species and Pythium species were investigated by using a Biolog$^{(R)}$ microplate assay to determine if there are differences in the carbon utilizations of selected strains of these species. It may be possible to afford a competitive advantage to bacterial biological control agents by providing them with a substrate that they can readily use as a carbon source, for example, in a seed coating formulation. Microplates, identified as SFP, SFN and YT were used to identify spore-forming bacteria, nonspore-forming bacteria, and yeast, respectively. Bacterial and mycelial suspensions were adjusted to turbidities of 0.10 to 0.11 at 600 nm. One hundred microliters of each of the bacterial and mycelial suspension were inoculated into each well of each of the three types of microplates. L-arabinose, D-galactose, D-melezitose and D-melibiose of the 147 carbohydrates tested were found to be utilized only by bacteria, and not by Pythium species, by Biolog$^{(R)}$ microplate assay, and this was confirmed by traditional shake flask culture. Thus, it indicated that the Biolog$^{(R)}$ microplate assay could be readily used to search for specific carbon sources that could be utilized to increase the abilities of bacterial biological control agents to adapt to contrived environments.

Microwave-assisted Protein Digestion on Various Locations of a Microplate

  • Lee, Ji-Hye;Park, Se-Hwan;Lee, Sun-Young;Hong, Jang-Mi;Park, Kyu-Hwan;Kim, Hyun-Sik;Kim, Jeong-Kwon
    • Mass Spectrometry Letters
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    • v.2 no.4
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    • pp.84-87
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    • 2011
  • The effectiveness of microwave-assisted protein digestion in different well positions of a 96-well microplate was investigated where microwave-assisted protein digestion of bovine serum albumin was performed in 10 different wells of a 96-well microplate in a microwave oven. Similarly increased sequence coverages (~70%) were generally observed for the 10 microwave-assisted protein digestion samples compared to conventional overnight digestion (63%), which is possibly due to higher miscleavage ratios (~53%) of the samples from microwave-assisted protein digestion than conventional overnight digestion (42.1%). The reproducible results of microwave-assisted digestions from different well positions demonstrate the potential of high-throughput analysis of proteins using microwave-assisted protein digestion.

Microplate Assay Measurement of Cytochrome P450-Carbon Monoxide Complexes

  • Choi, Suk-Jung;Kim, Mi-Ra;Kim, Sung-Il;Jeon, Joong-Kyun
    • BMB Reports
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    • v.36 no.3
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    • pp.332-335
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    • 2003
  • Cytochrome P450 in microsomes can be quantitated using the characteristic 450 nm absorption peak of the CO adduct of reduced cytochrome P450. We developed a simple microplate assay method that is superior to previous methods. Our method is less laborious, suitable for analyzing many samples, and less sensitive to sample aggregation. Microsome samples in microplate wells were incubated in a CO chamber rather than bubbled with CO gas, and then reduced with sodium hydrosulfite solution. This modification allowed a reliable and reproducible assay by effectively eliminating variations between estimations.

A Colorimetric Microplate Assay Method for High Throughput Analysis of Lipase Activity

  • Choi, Suk-Jung;Hwang, Jung-Min;Kim, Sung-Il
    • BMB Reports
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    • v.36 no.4
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    • pp.417-420
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    • 2003
  • The present work describes a colorimetric microplate assay for lipase activity based on the reaction between 5,5'-dithiobis(2-nitro benzoic acid) (DTNB) and the hydrolysis product of 2,3-dimercapto-1-propanol tributyrate (DMPTB). Reaction mixtures containing DTNB, DMPTB, and lipase were prepared in microplate wells, and the absorbance at 405nm was recorded after incubation at $37^{\circ}C$ for 30 min. A linear relationship was obtained in the range of 0.1-1 U of lipase activity by this method. The reaction conditions were also optimized for the range of 0.01-0.1 U or 1-10 U. When assaying crude tissue extracts, the reaction of DTNB with non-specific reducing agents created a major source of error. However, this error was corrected by the use of blank samples that did not contain DMPTB.

Screening of Hyaluronidase Inhibitory Activity Using a Microplate Assay (Microplate방법을 이용한 Hyaluronidase 저해 활성 검색)

  • Jeong, Sei-Joon;Kim, Na-Young;Ahn, Nyeon-Hyoung;Kim, Youn-Chul
    • Korean Journal of Pharmacognosy
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    • v.28 no.3
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    • pp.131-137
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    • 1997
  • The aqueous and methanolic extracts of 110 crude drugs were screened for hyaluronidase inhibitory activity using a microplate assay. Among them, MeOH extract of 15 crude drugs inhibited more than 80% of hyauluronidase activity at the concentration of 5mg/ml. The active principles of Anemarrhenae Rhizoma, Rhei Rhizoma, Ephedrae Herba, Pteropi Faeces and Ginseng Radix alba were transferred into organic solvents.

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Blood Folate Level Determined by a Microplate Reader and Folate Intake Measured by a Weighted Food Record (Microplate Reader를 이용하여 측정한 혈액의 엽산 농도와 실측량 기록법에 의한 엽산 섭취량)

  • 현태선;한영희;임은영
    • Korean Journal of Community Nutrition
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    • v.4 no.4
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    • pp.512-520
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    • 1999
  • Microbiological method using a 96-well microplate reader for folate assay was established, and folate intake and blood folate concentrations of 23 female college students were assessed. To evaluate folate intake, dietary data were collected by a 3-day weight food record, and serum and RBC folate concentrations were measured by the new method. The coefficient of variation for the new method was less than 10%. Mean daily folate intake of the subjects was 126.7${\mu}g$ which is only 50.7% of the RDA. The mean concentrations of serum and RBC folate were 7.46ng/ml and 294.4ng/ml, respectively, which were within the normal range. These results indicate that folate intake seems to be underestimated due to incomplete food composition database. Therefore, folate database should be appropriately in order to asses folate intake accurately.

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The Effect of $_L$=lysine on Growth Inhibition of Microcystis sp. ($_L$ =lysine에 의한 Microcystis sp.의 선택적 성장억제)

  • 송석환;신규철;한명수;최영길
    • Korean Journal of Environmental Biology
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    • v.21 no.2
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    • pp.216-221
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    • 2003
  • Various physico-chemical and biological methods have been used to remove. cyanobacteria which causes blooms and releases toxin. The purpose of the following experiment is aimed finding out which cyanobacteria are affected by $_L-lysine $ and what concentration of$_L-lysine $ inhibits cyanobacteria. The 20 samples of Microcystis sp. have been tested. To prove the growth inhibition on Microcystis sp., double-layered agar method and microplate method have been used. When the concentration of $_L-lysine $ is as heavy as 100 ${\mu}g\; ml^{-1}$~300 ${\mu}g\; ml^{-1}$, some Microcystis sp. have made halo zone. Some Microcystis sp. have shown so high activity as to be inhibited in their growth by the $_{L}$-lysine of concentration 10 ${\mu}g\; ml^{-1}$ with microplate method. These activities are various in accordance with every species. In additions, the microplate method has been proven to be an easy way which examine the lytic activity on the species of algae.e.

Antimycotic Susceptibility Testing of Trichophyton Rubrum by Microculture Method (96-well microplate를 이용한 Trichophyton Rubrum의 항진균제 감수성검사)

  • Lee, Moo-Woong;Kim, Jong-Chul;Choi, Jong-Soo;Kim, Ki-Hong
    • Journal of Yeungnam Medical Science
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    • v.9 no.2
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    • pp.396-406
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    • 1992
  • Various susceptibility tests have been used to determine minimal inhibition concentration(MIC) of dermatophytes. They have limitations to apply practically because they need long time to determine MIC. Authors examined MIC of T. rubrum to ketoconazole and itraconazole using 96-well microplate and 24-well macroplate by method of Granade and Artis and tried to check the possibility of this method on clinical application. Nine strains of T. rubrum from patients with dermatophytosis were used. Evaluations of the factors affecting MIC were also tried. The results were as follows. 1. Effect of inoculation density on determination time and MIC : Determination of MIC were possible in 4th days after inoculation at higher inoculation density Caborbance 2.0, 1.0) compared to 6th days at lower inoculation density(absorbance 0.5, 0.25). 2. Effect of incubation temperature on MIC : When incubating at $37^{\circ}C$, MIC were below 0.006-$0.04{\mu}g/ml$ to ketokckonazole and below 0.006-$0.04{\mu}g/ml$ to itraconazole while at $25^{\circ}C$ 0.08-$5.68{\mu}g/ml$ to ketoconazole and 0.006-$0.71{\mu}g/ml$ to itraconazole. Significant reduction of MIC was observed at $37^{\circ}C$ compared to $25^{\circ}C$. 3. Effect of container size on determination time and MIC : When incubating in 96-well microplate and 24-well macroplate, determination of MIC was possible in 4th to 6th days after inoculation in broth-containig 96-well microplate compared to 8th to 12th days in broth-containing 24-well macroplate. But no difference in MIC was observed between different container size. 4. Effect of media on MIC : When using broth as media, MIC were below 0.006-$5.68{\mu}g/ml$ to ketoconazole, below 0.006-$0.36{\mu}g/ml$ to itraconazole in broth-containg 24-well macroplate. When using agar as media, MIC were below 0.006-$5.68{\mu}g/ml$ to ketoconzole, below 0.006-$5.68{\mu}g/ml$ to intraconzole in agar-containing 24-well macroplate. There was slight increase of MIC with agar media compared to broth media. 5. These findings confirm that determination of MIC of dermatophtes by method of Granade and Artis is fast and simple technique for antifungal susceptibility test.

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A Rapid Method for Estimating the Levels of Urinary Thiobarbituric Acid Reactive Substances for Environmental Epidemiologic Survey

  • Kil, Han-Na;Eom, Sang-Yong;Park, Jung-Duck;Kawamoto, Toshihiro;Kim, Yong-Dae;Kim, Heon
    • Toxicological Research
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    • v.30 no.1
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    • pp.7-11
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    • 2014
  • Malondialdehyde (MDA), used as an oxidative stress marker, is commonly assayed by measuring the thiobarbituric acid reactive substances (TBARS) using HPLC, as an indicator of the MDA concentration. Since the HPLC method, though highly specific, is time-consuming and expensive, usually it is not suitable for the rapid test in large-scale environmental epidemiologic surveys. The purpose of this study is to develop a simple and rapid method for estimating TBARS levels by using a multiple regression equation that includes TBARS levels measured with a microplate reader as an independent variable. Twelve hour urine samples were obtained from 715 subjects. The concentration of TBARS was measured at three different wavelengths (fluorescence: ${\lambda}-_{ex}$ 530 nm and ${\lambda}-_{em}$ 550 nm; ${\lambda}-_{ex}$ 515 nm and ${\lambda}-_{em}$ 553 nm; and absorbance: 532 nm) using microplate reader as well as HPLC. 500 samples were used to develop a regression equation, and the remaining 215 samples were used to evaluate the validity of the regression analysis. The induced multiple regression equation is as follows: TBARS level (${\mu}M$) = -0.282 + 1.830 ${\times}$ (TBARS level measured with a microplate reader at the fluorescence wavelengths ${\lambda}-_{ex}$ 530 nm and ${\lambda}-_{em}$ 550 nm, ${\mu}M$) -0.685 ${\times}$ (TBARS level measured with a microplate reader at the fluorescence wavelengths ${\lambda}-_{ex}$ 515 nm and ${\lambda}-_{em}$ 553 nm, ${\mu}M$) + 0.035 ${\times}$ (TBARS level measured with a microplate reader at the absorbance wavelength 532 nm, ${\mu}M$). The estimated TBARS levels showed a better correlation with, and are closer to, the corresponding TBARS levels measured by HPLC compared to the values obtained by the microplate method. The TBARS estimation method reported here is simple and rapid, and that is generally in concordance with HPLC measurements. This method might be a useful tool for monitoring of urinary TBARS level in environmental epidemiologic surveys with large sample sizes.