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81.
Two fluorophore-nitrilotriacetic acid conjugates, Pro-Q Sapphire 365 and Pro-Q Sapphire 488 oligohistidine gel stains, have been developed for the fluorescence detection of fusion proteins containing oligohistidine tags directly in sodium dodecyl sulfate polyacrylamide gels, without the requirement for electroblotting, reporter enzymes or secondary detection reagents. Pro-Q Sapphire 365 oligohistidine gel stain exhibits bright-blue fluorescence (emission maximum = 450 nm) when illuminated with UV-A or UV-B light from a standard ultraviolet transilluminator. Pro-Q Sapphire 488 oligohistidine gel stain exhibits bright-green fluorescence (emission maximum = 515 nm) when illuminated with visible light from a laser-based gel scanner equipped with a 470 nm second-harmonic generation (SHG) or 488 nm argon-ion laser source. Typically, 25-65 ng of oligohistidine-tagged fusion protein in whole cell lysates is detectable using either stain. After documenting the fluorescence signal from the Pro-Q Sapphire dyes, gels may be post-stained with the red-fluorescent SYPRO Ruby protein gel stain in order to reveal the total protein pattern. 相似文献
82.
83.
Determination of organophosphate and carbamate pesticides based on enzyme inhibition using a pH-sensitive fluorescence probe 总被引:1,自引:0,他引:1
A flow injection system for the determination of organophosphate and carbamate pesticides is described. A sensitive fluorescence probe was synthesized and used as the pH indicator to detect the inhibition of the enzyme acetylcholinesterase (AChE). The percentage inhibition of enzyme activity is correlated to the pesticide concentration. Several parameters influencing the performance of the system are discussed. The detection limits of 3.5, 50, 12 and 25 μg/l for carbofuran, carbaryl, paraoxon and dichlorvos, in pure water, respectively were achieved with an incubation time of 10 min. A complete cycle of analysis, including incubation time, took 14 min. The detection system has been applied to the determination of carbofuran in spiked vegetable juices (Chinese cabbage and cole), achieving recovery values between 93.2 and 107% for Chinese cabbage juice and 108 and 118% for cole juice at the different concentration levels assayed. 相似文献
84.
血红蛋白过氧化物模拟酶胶束催化显色体系 总被引:6,自引:0,他引:6
在胶束Tween80介质中,研究了以血红蛋白(hemoglobin,Hb)作为过氧化物模拟酶、隐性亮绿(recessive brilliant green,RGB)为氢供体底物、溶解氧为受氢体的酶催化反应特性。在pH5.64(NH4)2HPO4-KH2PO4缓冲溶液中,利用模拟酶对溶解氧作为受氢体催化RBG氧化生成亮绿(brilliant green,BG)而拟定了测定Hb含量的新方法。讨论了胶束介质对酶体系催化反应的影响及酶催化反应的可能机理。 相似文献
85.
86.
Enhancement of electronic excitation energy transfer in the colloidal crystals of colloidal silica suspensions doped with fluorescent dyes 总被引:1,自引:0,他引:1
Kiyoshi Shibata Hiroshi Kimura Akira Tsuchida Tsuneo Okubo 《Colloid and polymer science》2006,285(2):127-133
The efficiency of electronic excitation energy transfer from photo-excited rhodamine 110 (Rh110, energy donor) to rhodamine B (RhB, energy acceptor) in an exhaustively deionized colloidal silica suspension has been studied. This colloidal suspension shows Bragg reflection due to the formation of colloidal crystals and the Bragg-peak wavelength is controllable by the volume fraction of the silica spheres. When the Bragg-peak wavelength matches with the fluorescence band of Rh110, a depletion was observed in the Rh110 fluorescence spectrum. This means the fluorescence of Rh110 is partially trapped due to the Bragg reflection inside the crystal lattice. In the coexistence of RhB, the enhancement of RhB fluorescence intensity was observed. These facts clearly indicate the trapped photon energy of Rh110 is efficiently transferred to RhB within the colloidal crystals. The quantitative measurements showed that the enhancement of the transfer efficiency is 20% (or slightly more) in the present experimental conditions. 相似文献
87.
在pH6.4的磷酸二氢钾-磷酸氢二钾缓冲液和CIMAJB、正丁醇、正庚烷组成的微乳液的介质中,水杨基荧光酮与铜(Ⅱ)、铬(Ⅵ)的显色体系灵敏度都有很大提高,选择了合适的测定波长,利用双波长解方程组法同时测定了底泥中的铜和铬。结果令人满意。 相似文献
88.
Hiroki Higuchi Hidetoshi Kawai Masakazu Ohkita Takanori Suzuki 《Tetrahedron letters》2004,45(15):3027-3030
The newly prepared tetracyanoanthraquinodimethane (TCNAQ) derivatives 1a,b with a chiral auxiliary are good electron acceptors and exhibit weak circular dichroism (CD) based on the absorption of TCNAQ. The twin-type electron acceptor 1c with two TCNAQ units shows larger ellipticity by exciton coupling. UV-vis, CD, and fluorescence spectra were changed drastically upon electrochemical reduction of 1c, which demonstrates the unprecedented three-way-output response system. 相似文献
89.
90.
Paraskevas D. Tzanavaras Demetrius G. Themelis Anastasios Economou Georgios Theodoridis 《Mikrochimica acta》2003,142(1-2):55-62
Two new simple and rapid methods are reported for the accurate and precise spectrophotometric determination of captopril
(CPL) using flow (FI) and sequential injection (SI) analysis. The methods are based on the fast oxidation of CPL by Fe(III).
The produced Fe(II) reacts with 2,2′-dipyridyl-2-pyridylhydrazone (DPPH) in acidic medium to form a colored complex which
is monitored spectrophotometrically at 535 nm. Both methods allow the determination of the analyte up to 1000 mg L−1 at a sampling rate of 120 and 60 injections per hour for FI and SI, respectively. The methods are very precise [s
r=0.8 and 1.2% at 500 mg L−1 CPL (n=12) for FI and SI, respectively] and the 3σ detection limits (c
L=4.0 and 7.0 mg L1, respectively) are quite satisfactory. Their application to a variety of anti-hypertensive commercial pharmaceutical formulations
showed excellent results (relative errors, e
r, < ± 1.6% in all cases compared to an official HPLC method), while common pharmaceutical excipients were found not to interfere.
Recovery experiments further verified the accuracy of the developed methods, as the percent recoveries were in the range of
98.1–102.5%.
Author for correspondence. E-mail: themelis@chem.auth.gr
Received May 9, 2002; accepted January 8, 2003
Published online May 5, 2003 相似文献