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21.
A fast and matrix assisted laser desorption/ionization-mass spectrometry (MALDI-MS) compatible protein staining method in one- and two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (1- and 2-D SDS-PAGE) is described. It is based on the counterion dye staining method that employs oppositely charged two dyes, zincon (ZC) and ethyl violet (EV) to form an ion-pair complex. The protocol, including fixing, staining and quick washing steps, can be completed in 1-1.5 h depending upon gel thickness. It has a sensitivity of 4-8 ng, comparable to that of colloidal Coomassie Brilliant Blue G (CBBG) staining with phosphoric acid in the staining solution. The counterion dye stain does not induce protein modifications that complicate interpretation of peptide mapping data from MS. Considering the speed, sensitivity and compatibility with MS, the counterion dye stain may be more practical than any other dye-based protein stains for routine proteomic researches. 相似文献
22.
A background-free, fast protein staining method in polyacrylamide gel electrophoresis using an acidic dye, zincon (ZC) and a basic dye, ethyl violet (EV) is described. It is based on the counterion dye staining technique that employs two oppositely charged dyes to form an ion-pair complex in staining solution. The selective binding of free dye molecules to proteins in acidic solution produces bluish violet-colored bands. It is a rapid and end-point staining procedure, involving only fixing and staining steps that are completed in 1-1.5 h. The detection limit of this method is 8-15 ng of protein that is comparable to the sensitivity of the colloidal Coomassie Brilliant Blue G (CBBG) stain. Due to its sensitivity and speed, this stain may be more practical than any other dye-based stains for routine laboratory purposes. 相似文献
23.
Characterization of amiodarone metabolites and impurities using liquid chromatography/atmospheric pressure chemical ionization mass spectrometry 总被引:1,自引:0,他引:1
Myung SW Chang YJ Min HK Kim DH Kim M Kang TK Yoo EA Sohn YT Yim YH 《Rapid communications in mass spectrometry : RCM》2000,14(21):2046-2054
Using the high performance liquid chromatography/atmospheric pressure chemical ionization tandem mass spectrometry (HPLC/APCI-MS/MS) technique, together with established trends from the literature, the structures of metabolites and impurities of amiodarone, an anti-arrhythmic drug, have been assigned. By comparing analyses of products of incubation with rat liver microsomes with controls in which glucose 6-phosphate dehydrogenase was omitted, metabolites could be distinguished from impurities. Structures for the two proposed metabolites and four impurities are proposed. 相似文献
24.
A radiochemical procedure is described for the fast and sensitive measurement of uranium isotopes in gaseous and liquid effluents of nuclear facilities. Equally, this procedure is suitable to measure uranium isotopes in all kinds of environmental samples. Uranium is leached from ashed sample materials with HNO3, HF, and Al(NO3)3 solution and separated from matrix elements by extraction with trioctylphosphinic oxide and backextraction with NH4F. After radiochemical cleaning by coprecipitation with LaF3 and anion exchange, uranium isotopes are electroplated on stainless steel discs from HCl/oxalate solution. The preparation is measured by alpha-spectrometry using surface barrier detectors. The detection limit for 1000 minutes of counting time is 2 mBq per sample and nuclide, the chemical yield is in the range of 50 to 80%. 相似文献
25.
Nam-Sook Kang Seok-Joo Hong Chong-Hak Chae Sung-Eun Yoo 《Journal of Molecular Structure》2007,820(1-3):58-64
A comparative molecular field analysis (CoMFA) for phosphodiesterase (PDE) IV inhibitors has been performed to correlate their chemical structures with their observed biological activity. In this study, CoMFA model based on docking mode for active site of PDE IV can describe the relative change in magnitude of the steric and electrostatic fields as a function of the compounds. Pyridine N-oxide and pyridine group of each compound are aligned toward the metal ion in S2-sub pocket of PDE IV. The study provided a statistically valid model with good correlation and predictive power, and consequently we identified some key features that may be used to design new derivatives. 相似文献
26.
27.
Jeong Hyun Kim Seung-Hyun Kang Il Nam Jung Kwan Mook Kim Bok Ryul Yoo Myong Euy Lee 《Journal of Sol-Gel Science and Technology》2004,32(1-3):31-35
Reaction of (triphenylmethyl)silanetriol (1) with cyclopentadienyltitanium trichloride (CpTiCl3) in the presence of triethylamine as a HCl scavenger gave both compounds of a partial open-cage type {[Ph3CSiO(OH)]3(Ph3CSiO3/2)(CpTiO3/2)4} (2) and cube type (Ph3CSiO3/2CpTiO3/2)4 (3). The 1:1 reaction of 1 and CpTiCl3 in toluene solvent at reflux temperature for 3 d afforded compounds 2 (22%) and 3 (36%). When 1 is reacted with a 1.5 fold excess of CpTiCl3 under the same conditions, compound 3 was obtained in high yield (81%) along with 2 in trace quantities. Compounds 2 and 3 were fully characterized by the analyses of 1H, 13C, 29Si NMR, IR, and FAB MS data. The solid-state structure of 3 was determined by a single crystal X-ray diffraction study. Compound 3 had shown to have catalytic activity for the oxidation of alkenes such as 1-octene, cyclooctene, and norbornene with t-butyl hydrogen peroxide. The effect of solvent was observed in this epoxidation reaction. The order of reactivity were decreased as follows: CHCl3 > hexane THF. 相似文献
28.
FTY720, a synthetic sphingoid base analog, was examined as a new sphingosine kinase inhibitor, which converts endogenous sphingosine into its phosphate form. With 20 microM of FTY720, sphingosine accumulated in the LLC-PK(1) cells in a time- and dose-dependent manner. The FTY720 treated cells showed a high concentration of fragmented DNA, a high caspase-3 like activity and TUNEL staining cells. It was also found that the sphingosine and sphinganine level increased in a time- and dose-dependent manner within 12 h after the FTY720 treatment. The sphingosine kinase activity was reduced by FTY720 as much as other sphingosine kinase inhibitors, N, N-dimethylsphingosine (DMS), dl-threo-dihydrosphingosine (DHS). The fragmented DNA content as a result of the 20 microM of FTY720 treatment and by 5 microM of the exogenously added BSA-sphingosine complex indicated typical apoptosis. Under similar conditions, the accumulated sphingosine concentration in all the cells was almost identical even though the sphingosine distribution inside the cells was somewhat different. These results indicate that the FTY720 induced apoptosis is associated with the inhibition of the sphingosine kinase activity and is strongly associated with the successive accumulation of sphingosine. 相似文献
29.
A one-step chromatographic method capable of separating all isomers of polyethylene glycol (PEG)-growth hormone-releasing factor (GRF) (1-29) conjugates was developed. The unmodified GRF (1-29) and seven different isomers of PEG-GRF (1-29) conjugates were separated by using a simple reversed-phase HPLC method depending on the differences of hydrophobicity due to the number and site of PEG attachment. The PEGylation sites of all isomers of PEG-GRF (1-29) conjugates were identified by determining the molecular masses of the Lys-C digested fragments with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. This study is a first report for the separation of all PEG-conjugate isomers and would be useful for further studies to find the promising conjugate by evaluating biological activity and stability of each isomer. 相似文献
30.
Park JI Lee WR Bae SS Kim YJ Yoo KH Cheon J Kim S 《The journal of physical chemistry. B》2005,109(27):13119-13123
In this paper, we describe an easy and reliable method for the production of patterned monolayers of Co nanoparticles. A two-dimensional monolayer of Co nanoparticles is fabricated by spreading a nanoparticle solution over an air-water interface and then transferring it to a hydrophobic substrate by using the Langmuir-Blodgett (LB) method. Transmission electron microscopy (TEM) was used to show that, with increasing surface pressure, the Co nanoparticles become well-organized into a Langmuir monolayer with a hexagonal close-packed structure. By controlling the pH of the subphase, it was found that a monolayer of Co nanoparticles with long-range order could be obtained. Further, by transferring the Langmuir monolayer onto a poly(dimethoxysilane) (PDMS) mold, the selective micropatterning of the Co nanoparticles could be achieved on a patterned electronic circuit. The electronic transport properties of the Co nanoparticles showed the ohmic I-V curve. 相似文献