• 제목/요약/키워드: fluorescence polarization method

검색결과 14건 처리시간 0.018초

QUANTITATION OF BARBITURATES IN URINE BY GC/MS AND ITS COMPARISON TO FLUORESCENCE POLARIZATION IMMUNOASSAY

  • Choo, Hea-Young;Park, Jeongeun;Park, Myung-Ja
    • Toxicological Research
    • /
    • 제7권1호
    • /
    • pp.29-35
    • /
    • 1991
  • Barbiturates are commonly abused tranquilizer and a rapid method to determine these drugs in biological samples is needed. In this study, was screened barbiturates in urine specimens by the fluorescence polarization immunoassay method(FPIA) and the positive samples were confirmed and identified by the more definitive GC/MS method. Fifteen positive smples which have barbiturate values higher than 0.5 ng/ml were analyzed by the GC/MS method. Eight samples were identified as phenobarbital and five samples were identified as crotilbarbitone.

  • PDF

Effect of iron on the proteolytic activity of live Uronema marinum (Ciliata: Scoticociliatida) measured by fluorescence polarization

  • 이은혜;권세련;최승혁;김기홍
    • 한국어병학회지
    • /
    • 제19권1호
    • /
    • pp.83-86
    • /
    • 2006
  • Effect of iron on the extracellular proteolytic activity of live Uronema marium was determined by fluorescence polarization (FP) method. Supplementation of 0.5 and 5.0 μM iron significantly increased caseinolytic activity of live U. marinum. In contrast, supplementation of 50 μM iron showed no significant differences in FP values compared to the control. The present result suggests that iron in cultured water or skin tissue of olive flounder may influence on the penetration and establishment of U. marinum, correlating with modulation of extracellular protease activity of the ciliates.

Label-Free and Real-Time Monitoring of Phosphatase Reactions Using a Phosphate-Specific and Fluorescent Probe

  • Lee, Ji-Hoon;Ahn, Hee-Chul;Shin, Dong-Yun;Ahn, Dae-Ro
    • Bulletin of the Korean Chemical Society
    • /
    • 제29권5호
    • /
    • pp.943-947
    • /
    • 2008
  • A phosphate-specific and fluorescent probe was prepared for label-free phosphatase assays based on fluorescence polarization. By using the probe, dephosphorylation reactions of DNA and protein substrates by calf intestinal alkaline phosphatase (CIP) could effectively be monitored in real-time. Since this assay method does not require additional materials such as labeled substrates and phosphospecific antibodies to obtain fluorescence polarization signals, it is simple, cost-effective, and expected to be useful not only for measuring activity of phosphatases but also for high-throughput screening of phosphatase inhibitors.

리포솜 지질막의 성질에 미치는 지질 조성의 영향 (Effects of Lipid Composition on the Properties of Phospholipid Liposomal Membranes)

  • 김민;한석규;김종국
    • 약학회지
    • /
    • 제38권2호
    • /
    • pp.131-139
    • /
    • 1994
  • Calcein-encapsulated small unilamellar vesicles of various lipid composition were prepared using the sonication technique, and their stabilities at $20^{\circ}C$ were examined by measuring calcein leakage from the liposomes. The fluidity of these liposomal bilayers was also investigated by measuring the fluorescence polarization of DPH labelled into the liposomes. The results showed that liposomes made of PC mixtures with different acyl chain length were very stable, which may be due to the formation of interdigitated bilayer structure. The addition of cholesterol further stabilized these PC liposomes. However, addition of cholesterol reduced the encapsulation efficiences of liposomes. The fluidity of the liposomes was significantly decreased by cholesterol in the liquid crystalline state, but not changed in the gel state. These results suggest that the enhanced stability of PC mixture liposomes may be ascribed to the formation of stable interdigitated bilayer structure. In membrane-mimetic and drug-delivery studies, vesicles made of mixtures of various phospholipids are recommended instead of addition of cholesterol to the phospholipid.

  • PDF

SCREENING OF BENZODIAZEPINES IN URINE BY THE IMMUNOASSAY AND QUANTITATION BY GC-NPD METHOD

  • Park, Jongsei;Park, Jeongeum;Park, Myung-Ja
    • Toxicological Research
    • /
    • 제7권1호
    • /
    • pp.21-27
    • /
    • 1991
  • We developed a simple method to determine benzodiazepines in biological samples using electron capature detectors and nitrogen-phosphorous detectors. The extraction of 13 benezodiazepines in urine at pH 9.5 with toluene and its analysis in GC/NPD showed the peaks in 9-16 min. In this retention time range, the biological backaground was fairly low and the drugs could be identified in low concentrations. The benzodiazepines in urine samples were screened by the fluorescence polarization immunoassay and positive samples were confirmed by the GC/NPD method.

  • PDF

Experimentally Measured Rotational Reorientation Time of Coumarin 6 Laser Dye in Ethanol and Acetonitrile Solvents

  • Renuka, C.G.;Raikar, U.S.
    • Journal of Photoscience
    • /
    • 제12권3호
    • /
    • pp.119-122
    • /
    • 2005
  • The photophysical properties of coumarin 6 laser dye have been studied in two solvents; ethanol and acetonitrile using steady-state fluorescence depolarization technique. The experimentally measured reorientation time of coumarin 6 is more or less the same in given solvents at particular temperature. It is found that coumarin 6 rotates slower in acetonitrile than in ethanol especially at higher values of viscosity over temperature. We also measure the ground and excited state dipole moments of coumarin 6 by solvent perturbation method. The results found that excited state dipole moment is greater than ground state dipole moment, which indicates that excited state is more polar than the ground state.

  • PDF

현미에서의 오크라톡신 A의 검색을 위한 형광편광면역분석법의 응용 (Application of Fluorescence Polarization Immunoassay for the Screening of Ochratoxin A in Unpolished Rice)

  • 박정현;정덕화;이인선
    • 생명과학회지
    • /
    • 제16권6호
    • /
    • pp.1006-1013
    • /
    • 2006
  • 식품안전에 대한 관심이 증가되고 있는 현재, 생물학적 화학적 위해요소로 분류되고 있고, 현재 많은 나라에서 규제치를 설정하고 있는 곰팡이 독소인 ochratoxin A(OTA)에 대한 정량적 측정이 가능한 고속검색법을 개발 하고자 단클론성 항체를 이용하여, 측정시 분리과정이 필요 없는 형광편광면역분석법(FPIA)을 개발하고 최적화 시켰다. 동일구조를 가지는 형광물질 표식자인 OTA-EDF를 합성하여 OTA에 대한 특이항체와 경쟁반응을 시켜 나타나는 형광-편광도(mP)의 변화를 측정하였다. 이는 면역분석법의 특이성과 민감성을 충분히 만족하였다. OTA의 검출범위는 0.5-200 ng/ml였고, 검출한계는 0.3 ng/ml였다. 개발된 분석법은 다른 곰팡이 독소들과의 교차반응은 없었고 높은 특이성과 재현성 및 회수율을 나타내었다. HPLC 방법에 의한 회수율은 88-84%로 다소낮게, FPlA법의 회수율은 90-110%로 다소 높게 나타났다. 16점의 현미시료를 분석하였을 때, 2점이 상관관계가 높게 12-20 ppb 정도 오염된 것으로 나타났다. 4점은 FPIA 및 HPLC 모두에서 음성으로 판정되었다. 개발된 FPIA는 복잡한 전처리 방법이 필요 없는 신속한 검색이 가능하므로, 식품 및 환경에서의 OTA 잔류 검사에 유용하게 사용될 수 있을 것이다.

Development of an Alignment Method for Retarders in isoSTED Microscopy

  • Ilkyu Park;Dong-Ryoung Lee
    • Current Optics and Photonics
    • /
    • 제8권4호
    • /
    • pp.421-426
    • /
    • 2024
  • The use of stimulated emission depletion (STED) microscopy has significantly improved resolution beyond the limits imposed by diffraction; Furthermore, STED microscopy adopts a 4Pi-geometry to achieve an isotropic improvement in resolution. In isoSTED microscopy, a polarizing beam splitter and retarders are used in a 4Pi cavity to split beams of identical power, generating constructive and destructive interference for lateral and axial resolution improvements, respectively. The precise alignment of the retarders is crucial for optimizing the performance of isoSTED microscopy, because this orientation affects the quality of the depletion focus, necessitating zero intensity at the center. Incomplete destructive interference can lead to unwanted fluorescence inhibition, resulting in degraded resolution and contrast. However, measuring the intensity and polarization state in each optical path of the 4Pi cavity is complex and requires additional devices such as a power meter. Here, we propose a simple and accurate alignment method for the 4Pi cavity in isoSTED microscopy. Our approach demonstrates the equal allocation of power between upper and lower beam paths and achieves complete destructive interference using a polarizing beam displacer and a single CCD camera positioned outside the 4Pi cavity.

Complex Detection Between Transcription Regulator and Promoter DNA by UV Spectroscopic Method

  • Lee, Kyungmin;Gang, Jongback
    • 통합자연과학논문집
    • /
    • 제5권3호
    • /
    • pp.163-167
    • /
    • 2012
  • UV spectrophotometer was used to detect protein-DNA complex from DNA melting profile under constant temperature increase. Melting temperature (Tm) was $43^{\circ}C$ in copA duplex DNA alone. In the presence of Proteus mirabilis transcription regulator protein (PMTR) protein at 0.2 and 0.4 ${\mu}M$, Tm's were $45{\pm}0.5$ and $47.6{\pm}0.6^{\circ}C$, respectively. According to fluorescence polarization and gel shift assay. PMTR:copA complex was detected by the retarded migration on gel and the dissociation constant ($K_d$) was $(9.2{\pm}2.8){\times}10^{-9}M$.

트란스-1,2-비스피라질에틸렌에 대한 분광학적 연구 (Spectroscopic Studies of trans-1,2-Bispyrazylethylene)

  • 심상철;강한영
    • 대한화학회지
    • /
    • 제22권4호
    • /
    • pp.239-244
    • /
    • 1978
  • Stilbene 유도체인 trans-1,2-bispyrazylethylene에 대한 분광학적 연구를 행하였다. 보통의 자외선 흡수스펙트럼에서는 $n{\rightarrow}{\pi}^*$ 흡수밴드는 에너지가 비슷한 ${\pi}{\rightarrow}{\pi}^*$ 흡수밴드에 가려져서 나타나지 않는데 2차 미분 스펙트럼과 저온(77˚K)에서의 스펙트럼으로 부터 $n{\rightarrow}{\pi}^*$ 흡수 밴드를 확인할 수가 있었다. 또한 ${\pi}{\rightarrow}{\pi}^*$ 흡수밴드들의 전이에너지를 PPP-SCF-CI MO 방법으로 계산한 결과 스펙트럼에서 얻은 값과 잘 일치했다. 형광스펙트럼, 형광편광스펙트럼, 그리고 PPP-SCF-CI MO 방법에 의한 계산으로부터 형광을 내는 상태는 $^1({\pi},\;{\pi}^*)$ 상태임을 알 수가 있었으며 이것은 형광에 대한 alkaline salt effect의 결과와 일치한다.

  • PDF