http://chineseinput.net/에서 pinyin(병음)방식으로 중국어를 변환할 수 있습니다.
변환된 중국어를 복사하여 사용하시면 됩니다.
96-well plate를 이용한 DPPH free radical 소거활성 측정과 그 응용
최정섭(Jung-Sup Choi),오정임(Jung-Im Oh),황인택(In-Tack Hwang),김성은(Sung-Eun Kim),전재철(Jae-Chul Chun),이병회(Byung-Hoi Lee),김진석(Jin-Seok Kim),김태준(Tae-Joon Kim),조광연(Kwang-Yun Cho) 한국농약과학회 2003 농약과학회지 Vol.7 No.2
A 96-well plate was applied to determine the DPPH free radical scavenging activity using 107 plant-specific enzyme inhibitors and 100 unknown plant-originated extracts. The final optimum volume was 250 ㎕ containing 100 μM DPPH ethanolic solution at pH 7.8. In this condition, the radical scavenging activities were significantly increased by two known antioxidants consisting of ascorbate and a-tocopherol in a concentration-dependent manner. Among the 107 inhibitors, ampicillin and gallic acid showed 90.2% and 92.6% antioxidant activity at 100 μM, respectively, and these results were consisted with previous findings. In the tested 100 natural materials at 50 ㎍/㎖, antioxidant activity of AT-407 resulted in the highest of 90.1%, and 10 extracts including AT-388 and AT-443 showed over 70%. Our results suggest that the use of 96-well plate for determining DPPH free radical scavenging activity would be a suitable method to select antioxidant-like substances of both synthetic compounds and natural products.
조미영,조재진,조태준,임정묵,이진 한국분자세포생물학회 2013 Molecules and cells Vol.35 No.5
Embryonic stem (ES) cells can be valuable for monitoring differentiation processes and for improving applications in basic developmental biology. The application of ES cells can be a useful tool for drug discovery and toxicology. Therefore, we suggest the high-throughput screening (HTS) system based on ES cells in this study. Firstly, we optimized the feeder-free condition and seeding cell number which can maintained for at least 7 days without over-con-fluency. We analyzed the system by cell viability, proliferation activity, RT-PCR and morphologic/immunohistochemical evaluations. The optimal cell seeding number was 30/well that was maintained the typical colonial morphology over 9 d with 1,000 U/ml LIF in the limited space. The cell in optimized condition expressed ALP, SSEA-1, Oct 4 and Nanog and the genetic expressions showed similar to protein expressions. The cell lineage marker expressions showed faint or none. The cell viability and proliferation activity were increased in time-dependent manner in our optimized HTS system. In conclusion, the novel HTS system using ES cells can by useful for developing models for drug discovery as well as toxicological screening in the near future.
Yuping Zhao,Xiaoqing Mu,Yao Nie,Yan Xu 한국식품과학회 2014 Food Science and Biotechnology Vol.23 No.6
A new, rapid spectrophotometric quantitativedetermination method for γ-decalactone (GDL) was developedbased on the ferric hydroxamate reaction. Samples addedto a mixture of NH2OH·HCl and NaOH were heated toreact. Then HCl was added. Ferric chloride was added forcoloration, and 75% (v/v) ethanol was added. Absorbancevalues were measured after coloring for 10 min. Theconcentration range of a GDL calibration curve was 0.125-8.14 g/L. A 96-well plate high-throughput screening methodwas established to obtain desirable strains. A total of 215GDL-producing strains were identified from 4,327 samples. The highest production was 179 mg/L.
Saccharomyces cerevisiae를 이용한 효율적인 호흡저해제 검정법
최경자(Gyung Ja Choi),김진철(Jin-Cheol Kim),김흥태(Heung Tae Kim),조광연(Kwang Yun Cho) 한국농약과학회 2000 농약과학회지 Vol.4 No.3
A rapid assay to determine respiration inhibition of Saccharomyces cerevisiae by chemicals was developed. S. cerevisiae was harvested with two different liquid media, yeast extract-pep tone-dextrose (YPD) medium capable of occurring both glucose fermentation and mitochondrial respiration, and non-fermentable carbon-yeast extract (NFY) medium capable of occurring respiration only. Wells in 96-well plate were loaded with each cell suspension and various concentrations of 46 fungicides with various modes of action. In NFY medium, the non-fermentable carbon source, ethanol (NFY-E medium), glycerol (NFY-G medium) or lactate (NFY-L medium), was used. After incubation for 1~3 days, minimum inhibitory concentrations (MICs) of the chemicals were recorded in the media. Of the 46 inhibitors employed in this study, four inhibitors of fungal respiration by blockage of electron flux in the mitochondrial respiratory chain, azoxystrobin, kresoxim-methyl, metominostrobin, and trifloxystrobin, exhibited strong antifungal activity in all of NFY media, but no activity in YPD medium. In contrast to this, five N-trihalomethylthio fungicides showed much stronger antifungal activities in YPD medium than three NFY media. Eleven fungicides inhibited growth of S. cerevisiae in all media and the other 26 fungicides showed no antifungal activity in all media. Thus, our rapid and efficient in vitro method can be considered as an alternative assay system for respiration inhibitor.
Acetolactate synthase에 대한 고효율 활성 측정방법 및 신규 저해제 탐색
박상희(S.H. Park),황인택(I.T. Hwang),이관휘(K.H. Lee),최정섭(J.S. Choi),변종영(J.Y. Pyon),조광연(K.Y. Cho) 한국농약과학회 2001 농약과학회지 Vol.5 No.3
This study was conducted to develop a high throughput system for screening acetolactate synthase (AlS) inhibitors, and to detect basic mother molecules for developing new novel herbicide candidates. The high throughput screening (HIS) method using 96-well plate and microplate reader was developed. This method is 8 times more effective than basic technique in one cycle per person. Futhermore, considering for less than 1/10 volume of materials required for ALS test and enzyme kinetics with 16 times faster speed compared to those of former procedure, this HIS method has more than 100 times higher efficacy than basic system in a consecutive procedure. We discovered 11 new ALS inhibitors such as 2-oxoglutaric acid, aminooxyacetic acid, azelaic acid, citric acid, cyanuric fluoride, itaconic acid, malonic acid, niclosamide, oxalic acid, glyoxylic acid, and suramin from 107 commercial plant-specific inhibitors using this technique. We hope these results might be useful to discover lead compounds for developing new novel herbicide candidate.
녹색반응을 이용한 클로로겐산의 함량측정을 위한 흡광도 분석법과 블루베리 잎에 함유된 클로로 겐산의 함량분석
Dong-Min Chung(정동민),Young Chul Chung(정영철),Hyo Kon Chun(전효곤) 한국생명과학회 2011 생명과학회지 Vol.21 No.4
본 연구는 클로로겐산의 함량을 측정하기 위한 흡광도 방법을 구축하였으며, 구체적으로는 클로로겐산이 50℃와 글라이신 및 알칼리 조건하에서 녹색반응이 일어난다는 현상을 이용했다. 녹색형성은 일련의 클로로겐산 농도에 의존성을 가졌다. 본 연구에 따른 클로로겐산의 측정방법을 이용하여 블루베리에 함유된 클로로겐산의 함량을 분석하였다(12.42 ㎎/g d.w). 이러한 방법은 저가의 비용과 빠른 시간으로 많은 시료를 대량으로 쉽게 클로로겐산의 함량은 측정할 수 있는 효과를 가진다. We developed a spectrophotometric assay for the quantitative determination of chlorogenic acid based on the formation of green pigment at 50℃ under glycine and alkaline conditions in 96-well plates. The formation of green pigment was linear with a series of chlorogenic acid concentration (0-300 μM). Using this method, the content of chlorogenic acid (12.42 ㎎/g dry weight) in the leaves of blueberry was quantified. This method is high-throughput, cost-effective, rapid, and easy to perform.