• Title/Summary/Keyword: fluorophore

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A Sensitive Fluorescent Detection Method for Prostaglandins by High Performance Liquid Chromatography (고속액체크로마토그라피에 의한 프로스타글란딘류의 고감도 형광 분석법)

  • Lee, Yong-Moon;Moon, Dong-Cheul
    • YAKHAK HOEJI
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    • v.36 no.5
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    • pp.506-511
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    • 1992
  • The Prostaglandins were derivatized rapidly with monodansyl cadaverine as a fluorophore in mild conditions. The carboxylic moiety of prostaglandins was activated with diethyl phosphorocyanidate and successively coupled with fluorophore in dimethylformamide at room temperature. The labeling yield was reached about 95% at 15 min using arachidic acid $(C_{20:0})$ as a test sample. This derivative showed constant fluorescent intensity at $4^{\circ}C$ for 180 days. The derivatives of prostaglandins were shown high solvent selectivity with tetrahydrofuran in reversed-phase column. therefore, these derivatives could be successfully separated on YMC pack A-212(S-5 120A C8) column in tetrahydrofuran-based eluents. The detection limits of these derivatives was ca. 500 fmol and determination limits was ca. 5 pmol as injected amount in fluorescent detection $({\lambda}ex.\;340\;nm,\;{\lambda}em.\;520\;nm)$. In this method, the ranges of recovery and coefficient of variation were $93.6{\sim}102.7%$ and $4.3{\sim}5.8%$, respectively.

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Utilizing Natural and Engineered Peroxiredoxins As Intracellular Peroxide Reporters

  • Laer, Koen Van;Dick, Tobias P.
    • Molecules and Cells
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    • v.39 no.1
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    • pp.46-52
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    • 2016
  • It is increasingly apparent that nature evolved peroxiredoxins not only as $H_2O_2$ scavengers but also as highly sensitive $H_2O_2$ sensors and signal transducers. Here we ask whether the $H_2O_2$ sensing role of Prx can be exploited to develop probes that allow to monitor intracellular $H_2O_2$ levels with unprecedented sensitivity. Indeed, simple gel shift assays visualizing the oxidation of endogenous 2-Cys peroxiredoxins have already been used to detect subtle changes in intracellular $H_2O_2$ concentration. The challenge however is to create a genetically encoded probe that offers real-time measurements of $H_2O_2$ levels in intact cells via the Prx oxidation state. We discuss potential design strategies for Prx-based probes based on either the redoxsensitive fluorophore roGFP or the conformation-sensitive fluorophore cpYFP. Furthermore, we outline the structural and chemical complexities which need to be addressed when using Prx as a sensing moiety for $H_2O_2$ probes. We suggest experimental strategies to investigate the influence of these complexities on probe behavior. In doing so, we hope to stimulate the development of Prx-based probes which may spearhead the further study of cellular $H_2O_2$ homeostasis and Prx signaling.

Modification of Carboxyl Residues of Proteins with Pyridoxamine as a Fluorophore

  • Kwon, Oh-Shin
    • BMB Reports
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    • v.29 no.3
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    • pp.215-220
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    • 1996
  • A general procedure to quantitate the reaction of carbodiimides with carboxy groups of proteins is described. Pyridoxamine reacts with the o-acylisourea intermediate generated during the reaction of carboxyl residues with carbodiimides. The extent of the reaction is determined by measuring the spectroscopic properties, absorption and emission, of pyridoxyl residues covalently attached to the proteins. Resolved pig brain aspartate aminotransferase (apoenzyme), inactivated by 1-ethyl-3-(3-dimethylamino propyl) carbodiimide, reacts with $[^{3}H]pyridoxamine$. After trypsin digestion, one peptide labeled with radioactive pyridoxyl was separated by reverse phase HPLC.

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TCE, PCE 측정을 위한 바이오센서의 특성

  • 류두현;김용미;최상일
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2004.04a
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    • pp.388-391
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    • 2004
  • A sol-gel fiber-optic biosensor with encapsulated pH-sensitive fluorophore and immobilized genetically modified toluene-o-xylene monooxygenase was developed to detect TCE and PCE, which are carcinogenic chlorinate organic compounds prevailing in ground water. The sensitivity was characterized for the composition of sol-gel, and manufacturing procedure. The intensity curve reveals a linear range of intensity for pollutant concentration range of 0.01ppm and 1ppm. The change in intensity was appeared to be larger at each of L for same condition, and, therefore, the wavelength of λ was chosen for the analytical measurement.

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Photo-induced Electron Transfer(PET) Based Luminescent Chemosensors Detecting Hazardous Substances (광유발 전자전이를 이용한 유해물질 인식 형광 화학센서의 개발)

  • Yoon, Sung-Ho
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.25 no.9
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    • pp.711-716
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    • 2012
  • Based on both organic synthesis and theoretical calculations on the effects of molecular orbital energy levels of amines on the fluorescence properties of the fluorophore, fluorescent "turn-on" chemosensors detecting hazardous substances, including aldehyde chemicals and $Hg^{2+}$ ion, were developed.

Shuffled toluene-o-xylene monooxygenase를 이용한 TCE 측정용 fluorescence biosensor

  • ;;;;Thomas Wood
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2003.09a
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    • pp.475-478
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    • 2003
  • A sol-gel fiber-optic biosensor with encapsulated pH-sensitive fluorophore and immobilized genetically modified toluene-o-xylene monooxygenase was developed to detect TCE, which is carcinogenic chlorinate organic compounds prevailing in ground water. The sensitivity was characterized for the composition of sol-gel, and manufacturing procedure.

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Synthesis and Characterization of Red Organic Fluorescent of Perylene Bisimide Derivatives (Perylene Bisimide 유도체의 적색 유기 형광체 합성 및 특성 연구)

  • Lee, Seung Min;Jeong, Yeon Tae
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.30 no.9
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    • pp.577-582
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    • 2017
  • The white light of a hybrid LED is obtained by using red and green organic fluorescent layers made of polymethylmethacrylate (PMMA) films, which function as color down-conversion layers of blue light-emitting diodes. In this research, we studied the fluorescence properties of a red organic fluorophore, employing perylene bisimide derivatives applicable to hybrid LEDs. The solubility, thermal stability, and luminous efficiency are important characteristics of organic fluorophores for use in hybrid LEDs. The perylene fluorescent compounds (1A and 1B) were prepared by the reaction of 4-bromophenol and 4-iodophenol with N,N'-bis(4-bromo-2,6-diisopropylphenyl)-1, 6,7,12-tetrachloroperylene-3,4,9,10-tetracarboxyl diimide (1) in the presence of dimethyl formaldehyde (DMF) at $70^{\circ}C$. The synthesized derivatives were characterized by using $^1H-NMR$, FT-IR, UV/Vis absorption and PL spectra, and TGA analysis. Compounds 1A and 1B showed absorption and emission at 570 nm and 604 nm in the UV/Vis spectrum. We also documented favorable solubility and thermal stability characteristics of the perylene fluorophores in our work. Perylene fluorophore 1, with the 4-bromophenol substituent 1A, exhibited particularly good thermal stability and solubility in organic solvents.

Rapid Measurement of $NH_3$ and Weak Acid Permeation Through Liposomes and Renal Proximal Tubule Membranes

  • Bae, Hae-Rahn;Suh, Duck-Joon;Lee, Sang-Ho
    • The Korean Journal of Physiology
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    • v.28 no.2
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    • pp.203-214
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    • 1994
  • Using the methods of stopped-flow and epifluorescence microscopy with entrapped fluorophore, membrane permeability of $NH_3$ and weak acids in liposomes, renal brush border (BBMV) and basolateral membrane vesicles (BLMV), and primary culture cells from renal proximal tubule was measured. Permeability coefficient (cm/sec) of $NH_3$ was $(2.9{\times}10^{-2}$ in phosphatidylcholine liposome $25^{\circ}C)$, $5.9{\times}10^{-2}$ in renal proximal tubule cell $(37^{\circ}C)$, $4.0{\times}10^{-2}\;and\;2.4{\times}10^{-2}$ in BBMV and BLMV $(25^{\circ}C)$, respectively. Formic acid has the highest permeability coefficient among the weak acids tested, which was $4.9{\times}10^{-3}$ in liposome, $5.0{\times}10^{-3}$ in renal proximal tubule cell, $9.1{\times}10^{-3}$ in BBMV and $3.8{\times}10^{-3}$ in BLMV. There was a linear relationship between external concentration of nonionized formic acid and initial rate of flux of formic acid in liposome, and the slope coincided with the value of permeability coefficient of formic acid measured in pH 7.0. These results show that techniques of stopped-flow and epifluorescence microscopy with entrapped fluorophore provide the precise method of measurement of very rapid transport of nonelectrolytes through membranes with the advantages of instantaneous mixing effect, good resolution time and easy manipulation.

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