• 제목/요약/키워드: Fluorescent probe technique

검색결과 29건 처리시간 0.029초

Ibuprofenlysine binding to human and bovine serum albumin using a fluorescence probe technique

  • Kim, Chong-Kook;Cha, Hyun-Sook;Kim, Yang-Bae;Yu, Byung-Sul
    • Archives of Pharmacal Research
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    • 제4권1호
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    • pp.19-24
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    • 1981
  • The possibility of using a fluorescence probe technique for the study of ibuprofenlysine binding to human and bovine serum albumin was investigated. 1-anilino-8-naphalenesulfonate was used as the probe. The number of binding sites of human and bovine serum albumins for ibuprofenlysine appears to be 4 and 2, respectively. By using this technique, the association constants were found to be $1.533{\times}10^{4}M^{-1}$ and $2.238{\times}10^{4}M^{-1}$, respectively.

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The Effect of Cure History on the Fluorescence Behavior of an Unsaturated Polyester Resin with A Fluorescence Probe

  • Donghwan Cho;Yun, Suk-Hyang;Bang, Dae-Suk;Park, Il-Hyun
    • Macromolecular Research
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    • 제12권3호
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    • pp.282-289
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    • 2004
  • We have extensively characterized the fluorescence behavior of unsaturated polyester (UP) resin in the absence and presence of a 1,3-bis-(l-pyrenyl)propane (BPP) fluorescent probe at various dynamic and isothermal cure histories by means of a steady-state fluorescence technique using a front-face illumination equipment. In addition, we explored the effect of the fluorescence intensity on the relaxation of the fluorescent probe in the UP resin by resting the dynamically and isothermally cured resin at ambient temperature and pressure for 24 h. The monomer fluorescence intensity, which has two characteristic peaks at 376 and 396nm, changed noticeably depending on the cure temperature and time and provided important information with respect to the molecular and photophysical responses upon curing. The result of the fluorescence study indicates that the increased local viscosity and restricted molecular mobility of the UP resin surrounding the BPP probe after curing are both responsible for the enhancement of the monomer fluorescence intensity. Our results also demonstrate that once the BPP probe has enough time to rearrange and become isolated prior to fluorescence, a sufficient amount of fluorescence is emitted. Therefore, we note that the fluorescence behavior of this UP resin system is influenced strongly by the relaxation process of the fluorescent probe in the resin as well as process used to cure the resin.

Barbiturates가 생체세포막 외측 단층의 소수성 부위와 친수성 부위에 분포되는 상대적 비율 (The Region of Distribution of Barbiturates in Synaptosomal Plasma Membrane Vesicles Isolated from Rat Brain as Studied by Fluorescence Quenching)

  • 윤일;이병우
    • 대한약리학회지
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    • 제31권3호
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    • pp.271-278
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    • 1995
  • Barbiturates의 분자적 약리학적 작용기전 연구에 기초자료를 제공키 위하여 본연구를 수행하였다. 분자적 약리작용 기전 연구에서는 무엇보다도 선행되어야 하는 것이 barbiturates가 신경세포막에서 어느 부위에 주로 분포되는가를 알아내는 데에 있다. 쥐(Rat)의 뇌로부터 분리한 synaptosomal plasma membrane vesicles (RSPMV)를 분리한 후 이 RSPMV 외측 단층(outer monolayer)의 소수성 부위와 친수성 부위에 barbiturates가 분포되는 경향을 형광 probe 법으로 검색하였다. 세포막 외측 단층의 친수성 부위에 분포되는 형광 probe N-octadecylnaphthyl-2-amino-6-sulfonic acid (ONS)와 소수성 부위에 분포되는 형광 probe12-(9-anthroyloxy)stearic acid (AS)를 각각 봉입한 후 형광소광법으로 barbiturates의 분포를 측정한 결과는 다음과 같다. 1) 대부분의 barbiturates가 RSPMV 외측 단층의 친수성 부위(표면)에 분포되고 소수성 부위 (hydrophobic region)에 극히 소량만이 분포된다는 것을 확인하였다. 2) 마취효과를 크게 일으키는 barbiturates일수록 소수성 부위에 분포되는 양이 증가하였다. barbiturates 종류에 따른 RSPMV 외측 단층 소수성 부위에의 분포 크기는 thiopental sodium > pentobarbital > hexobarbital > amobarbital > phenobarbital의 순위였다.

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Bacillus 속 세균을 검출하기 위한 Fluorescent In Situ Hybridization 방법의 개발 (Development of FISH Methods for Detection of Genus Bacillus)

  • 홍선희;김옥선;송홍규;이동훈;안태석
    • 미생물학회지
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    • 제37권3호
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    • pp.204-208
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    • 2001
  • Bacillus속에 속하는 세균을 FISH 방법으로 검출하기 위한 새로운 방법을 개발하였다. LGC353b, S-G-Bacill-0597-a-A-22와 S-G-Bacill-0597-a-S-22 등 3개의 probe를 22종의 Bacillus속 세균의 혼합배양액에 적용한 결과 S-G-Bacill-0597-a-A-22의 probe가 DAPI로 검출된 세균 중 95%가 FISH로도 측정되어 가장 효율이 좋은 것으로 나타났다. Hybrillization 과정 전에 Bacillus의 영양세포와 포자의 세포벽을 파괴하기 위하여 SDS/DTT (dithiothreitol)와 lysozyme를 순차적으로 전처리하는 방법과 lysozyme용액으로만 전처리 하는 두 가지 방법을 사용하였다. 그 결과, Bacillus를 검출하기 위해서는 영양세포의 경우에는 lysozyme용액으로만 전처리 하는 방법이 가장 좋았으며, 포자는 FISH로 검출되지 않았다. 따라서 토양, 하수 처리장등의 생태계에서 Bacillus를 검출할 경우에는 영양세포는 lysozyme으로 전처리 하여 FISH로 검출하고 포자는 따로 포자염색으로 검출하는 두가지 방법을 병행하여야한다.

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n-Alkanols가 소의 대뇌피질로부터 분리한 Synaptosomal Plasma Membrane Vesicles의 측방확산운동 범위와 속도에 미치는 영향 (The Effect of n-Alkanols on the Lateral Diffusion of Synaptosomal Plasma Membrane Vesicles Isolated from Bovine Cerebral Cortex)

  • 정인교;강정숙;윤일
    • 대한약리학회지
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    • 제29권1호
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    • pp.157-163
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    • 1993
  • n-Alkanols의 분자적 약리작용기전 탐구에 기초자료를 제공하기 위하여 소의 신선한 대뇌피질로부터 분리한 synaptosomal plasma membrane vesicles (SPMV) 지질 이중층의 측방확산운동에 미치는 n-alkanols의 영향을 형광 probe법으로 검색하였다. n-Alkanols는 SPMV 지질 이중층의 측방확산운동 범위와 속도를 농도 의존적으로 증가시켰고 1-nonanol까지는 탄소수가 두개 증가될 때마다 그 효력은 약 10배 가량 증가되었으나 탄소수 10개인 1-decanol의 효력은 오히려 감소되는 경향을 나타내었다.

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Fluorescent In Situ Hybridization방법으로 분석한 소양호 세균 군집 구조의 계절적 변화 (Seasonal Changes of bacterial community analysed by fluorescent in situ hybridization method in Lake Soyang)

  • 홍선희;안태석
    • 미생물학회지
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    • 제34권3호
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    • pp.169-174
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    • 1998
  • 소양호에서 세균군집의 계절적, 수심별 변화를 파악하고자 총세균수와 EUB338, ALG1b, BET42a, GAM42a와 CF probe 등 fluorescent rRNA target oligonuvleotide probe와 반응하는 세균 개체수를 측정하였다. 총세균수는 $0.5{\sim}2.01{\times}10^6cells{\cdot}ml^{-1}$이였으며, 2 m와 5 m 수층에서 높게 나타났다. 총세균수에 대한 Eubacteria의 비율은 22~100%이였고, Proteobacteria ${\alpha}$-group은 Eubacteria의 2.6~66.7%, ${\beta}$-group은 4.5~53.5%, ${\gamma}$-group은 4.6~76.7%, 그리고 Cytophage-Flavobacterium group은 2.1~35.9%이였다. 또한 세균군집은 계절별, 수심별로 다양한 변화를 보여, 겨울철은 ${\beta}$-group이, 봄철과 초여름은 ${\gamma}$-group이, 여름철은 ${\alpha}$-group이 우점하였고, Cytophage-Flavobacterium group이 특정적으로 우점하는 시기는 없었다. 이러한 세균 군집 구조의 분포로 계절별, 수심별로 호수에 대한 독특한 특징을 알 수 있었다.

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Characterization of the Surface Contribution to Fluorescence Correlation Spectroscopy Measurements

  • Chowdhury, Salina A.;Lim, Man-Ho
    • Bulletin of the Korean Chemical Society
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    • 제32권2호
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    • pp.583-589
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    • 2011
  • Fluorescence correlation spectroscopy (FCS) is a sophisticated and an accurate analytical technique used to study the diffusion of molecules in a solution at the single-molecule level. FCS is strongly affected by many factors such as the stability of the excitation power, photochemical processes, mismatch between the refractive indices, and variations in the cover glass thickness. We have studied FCS near the surface of a cover glass by using rhodamine 123 as a fluorescent probe and have observed that the surface has a strong influence on the measurements. The temporal autocorrelation of FCS decays with two characteristic times when the confocal detection volume is positioned near the surface of the cover glass. As the position of the detection volume is moved away from the surface, the FCS autocorrelation becomes one-component decaying; the characteristic time of the decay is the same as the faster-decaying component in the FCS autocorrelation near the surface. This observation suggests that the faster component can be attributed to the free diffusion of the probe molecules in the solution, while the slow component has its origin from the interaction between the probe molecules and the surface. We have characterized the surface contribution to the FCS measurements near the surface by changing the position of the detection volume relative to the surface. The influence of the surface on the diffusion of the probe molecules was monitored by changing the chemical properties of the surface. The surface contribution to the temporal autocorrelation of the FCS strongly depends on the chemical nature of the surface. The hydrophobicity of the surface is a major factor determining the surface influence on the free diffusion of the probe molecules near the surface.

아연결핍된 단핵구 U937 Cell Line에 있어서의 유전자 발현 탐색 : cDNA Microarray 기법 이용 (Gene Expression in Zn-deficient U937 Cell Line : Using cDNA Microarray)

  • Beattie, John H.;Trayhurn, Paul
    • Journal of Nutrition and Health
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    • 제35권10호
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    • pp.1053-1059
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    • 2002
  • In post-genome period, the technique for identifying gene expression has been changed to high throughput screening. In the field of molecular nutrition, the need for this technique to clarify molecular function of the specific nutrient is essential. In this study, we have tested the zinc-regulated gene expression in zinc-deficient U937 cells, using cDNA microarray which is the cutting-edge technique to screen large numbers of gene expression simultaneously. The study result can be used for the preliminary gene screening data for clarifying, using monocyte U937 cell line, molecular Zn aspect in atherosclerosis. U937 cells were cultured in Zn-adequate (control, 12 $\mu$M Zn) or Zn-deficient (experimental, 0 $\mu$M Zn) ESMI media during 2 days, respectively. Cells were harvested and RNA was extracted. Total RNA was reverse-transcriptinized and synthesized cDNA probe labeled with Cy-3. fluorescent labeled cDNA probe was applied to microarray slide for hybridization slide, and after then, the slide was scanned using fluorescence scanner. ‘Highly expressed genes’ in Zn-deficient U937 cells, comparing to Zn-adequate group, are mainly about the genes for motility protein, immune system protein, oncogene and tumor suppressor and ‘Less highly expressed genes’ are about the genes for transcription, apoptosis associated protein, cell cycle, and several basic transcription factors. The results of this preliminary study imply the effectiveness of cDNA microarray for expression profiling of a singly nutrient deficiency, specially Zn. Furthur study, using tailored-cDNA array and capillary endothelial cell lines, would be beneficial to clarify molecular Zn function, more in detail.

The Distribution of Barbiturates in Model Membranes of Total Lipids and Total Phospholipids Extracted from Brain Membranes

  • Park, Chang-Sik;Lee, Seong-Moon;Chung, In-Kyo;Kim, Jin-Bom;Son, Woo-Sung;Jang, Hye-Ock;Yun, Il
    • BMB Reports
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    • 제33권3호
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    • pp.221-227
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    • 2000
  • The distribution of barbiturates in the model membranes of total lipids (SPMVTL) and total phospholipids (SPMVPL) extracted from synaptosomal plasma membrane vesicles was determined by employing a fluorescent probe technique. The two fluorescent probes 2-(9-anthroyl)stearic acid and 12-(9-anthroyl)stearic acid were utilized as probes for the surface and the hydrocarbon interior of the outer monolayer of the SPMVTL and SPMVPL, respectively. The Stern-Volmer equation of fluorescent quenching was modified to calculate the relative distribution. The analysis of preferential quenching of these probes by barbiturates indicates that pentobarbital, hexobarbital, amobarbital and phenobarbital are predominantly distributed on the surface area, while thiopental sodium has an accessibility to the hydrocarbon interior of the outer monolayer of the SPMVTL and SPMVPL. From these results, it is strongly suggested that the more effective penetration into the hydrocarbon interior of the outer monolayer of the membrane lipid bilayer could result in a higher general anesthetic activity.

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Assembly and electrical property of GFP/Cytochrome b562 Fusion Protein ontothe Au Substrate

  • 정성철;최정우;이원홍
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2003년도 생물공학의 동향(XII)
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    • pp.630-633
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    • 2003
  • Transfer of an electron from one site to another in a molecular or between molecules and/or electrodes is one of the most fundamental and ubiquitous processes in chemistry, biology and physics. In this study fusion proteins composed by green fluorescent protein(GFP) and cytochrome b562 were used in fabricating molecular array as an electron sensitizer and electron acceptor, Protein formation onto the substrate was performed by the self-assembly technique. The fusion protein film were analyzed using scanning probe microscope(SPM), Surface Plasmon Resornance(SPR) and hybrid STM/I-V. The results suggest that the proposed molecular photodiode can be used as a basic unit of the memory device.

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