999 resultados para Fluorescence polarization immunoassay (FPIA)


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The sensitized fluorescence intensity of terbium ion can be notably enhanced when the Tb3+-fleroxacin complex is exposed to 365 nm light. By the measurements of fluorescence spectra, phosphorescence spectra, fluorescence quantum yield and fluorescence lifetime of the system, it is proved that irradiation makes the complex undergo a photochemical reaction and produces a new terbium complex which is more favorable to intramolecular energy transfer. The mechanism of the photochemical fluorescence enhancement was discussed.

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A surface plasmon resonance biosensor has been used to determine antibody activity in serum. As a model system, the interaction of mouse IgG and sheep anti-mouse IgG polyclonal antibody was investigated in real time. The factors, including pH value, ionic strength, protein concentration, influencing electrostatic adsorption of mouse IgG protein onto carboxylated dextran-coated sensor chip surface, were studied. The procedures of mouse IgG protein immobilization and immune reaction were monitored in real time. The regeneration effect using the different elution reagents was also investigated. The same mouse IgG immobilized surface can be used for 100 cycles of binding and elution with only 0.38% loss per regeneration in reactivity. The results show that the surface plasmon resonance biosensor is a rapid, simple, sensitive, accurate and reliable detection technique for real-time immunoassay of antibody activity. The assay allows antibodies to be detected and studied in their native form without any purification. (C) 2000 Elsevier Science B.V. All rights reserved.

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Octadecylamine was derivatized with dansyl chloride (5-dimethylaminonaphthalene-1-sulfonyl chloride) In order to simplify and understand the LB films of fluorescent probe labeling proteins. its monolayer and multilayers in the absence and presence of stearic acid were deposited by LB technique. Fluorescence spectra and lifetimes of the fluorescent products were studied to elucidate the microenvironment of molecules in the LB films.

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The general and synchronous spectra of phospholipase A(2) (PLA(2)) isolated from Chinese agkistrodon blomhoffii Ussurensis snake venom were studied. The chromophores of PLA(2) were mainly contributed by tyrosine and tryptophane residues when the intervals between the excitation wavelength and the emssion waveleagth (Delta lambda) were 20nm and 75nm, respectively. The pH of buffers could change the fluorescence intensities of PLA(2) by changing the charge distribution of its amino acid chain. Ca2+ can not only increase the emission fluorescence intensity of PLA(2) but also improve the reaction rate of PLA(2) with its corresponding substrate DPPC.

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The synchronous fluorescence spectra of myoglobin were studies for the first time. The fluorescence peals observed in the spectra were assigned, When the wavelength interval (Delta lambda) is 80 nm, the main peak at 335 nm is originated from the tryptophan residues in the myoglobin molecule. When Delta lambda is 20 nn, the peak at 308 nm is mainly due to the tyrosine residues in the myoglobin molecule and in a small part due to the tryptophan residues. Two peaks at 322 and 596 nm were observed in the spectrum of myoglobin for Delta lambda = 40 nm. The peak at 322 nm is due to both tyrosine and tryptophan residues. The peak at 596 nm is attributed to the heme group in the myoglobin molecule.

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Two new Ru(phen)(3)(2+)-based surfactants, Ru(phen)(2)(phenNHCO-C-11)(PF6)(2) and Ru(phen)(2)(phenNHCO-C-17)(PF6)(2), have been designed and synthesized, whose chemical structures were characterized by means of IR, H-1 NMR and MS. Also, electrochemistry and fluorescence of them are reported.

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Hemorrhagin III (AaH III) was separated and purified from the crude snake venom of Agkistrodon acutus, and its molecule weight was determined accurately to be 23; 284.4 +/- 0.1 by LDI1700-MALDI-TOF-MS. Emission spectra of AaH III showed that Trp residues were located by a great degree in the hydrophobic area. Addition of SDS and guanidine-HCl led to change of the molecular conformation of AaH III, and caused the fluorescence quenching of Trp residues. The red-shifted emission band of AaH III after adding guanidine-HCl showed that Trp residues exposed in polar solvents. The effects of pH, EDTA and metal ions on the fluorescence spectra of AaH III were also investigated.

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Four typical LB monolayer film materials, Ru(phen)(3)(2+) complexes with one ligand attached to different long chain alkyl amides, were designed and synthesized. Their chemical structures were identified by the techniques of FT-IR, H-1 NMR and ESI-MS. Also, UV-Vis, electrochemistry and fluorescence of these complexes are reported.

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A new series of Ru(II) polypyridine surfactants, Ru(bpy)(2)[phenNH CO(CH2)(n)CH3](PF6)(2), where n is 10, 12, 14 or 16, has been prepared and characterized. The H-1-NMR spectra of these new Ru(II) complexes were attributed and discussed by the H-1-H-1 COSY method. The comparative analysis of IR spectra of these complexes and their responding free ligands has shown that there are intensive d --> pi* feedback interactions between central Ru(II) and their ligands. Their fluorescent, electrochemical and electrochemiluminescent behaviours are also reported.

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Because of the extremely sensitivity to the local environment of the D-5(0) --> F-7(2) transition of Eu3+ ion, the fluorescence of Eu3+ ions was Studied by introducing Eu3+ ions to TiO2 gel by the sol-gel method, from which the structural changes of TiO2 gel were characterized. The results showed that the intensity of D-5(0) --> F-7(2) transition increased with the increasement of heat treatment temperature, which indicated the evaporation of molecular water and the completeness of the condensation reaction. Because of the quenching of the fluorescence induced by the cluster of Eu3+ ions, the addition of Al3+ ions greatly enhanced the emission intensity of Eu3+ ion.

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Ternary europium complex with dibenzoylmethane (DBM) and 1,10-phenanthroline (phen) was in-situ synthesized in thin SiO2/polyvinyl butyral (PVB) hybrid films by a two-step sol-gel process and characterized by;means of fluorescence spectroscopy. The luminescence spectra, fluorescence lifetimes and photostability were all investigated. The results showed that the hybrid films exhibited the characteristic emission bands of the Central rare earth Eu3+. In addition, Eu3+ presented longer fluorescence lifetime than in an ethanol solution and the complex had a higher photostability in the hybrid film than in the PVB film containing the corresponding pure complex.

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Fluorescence of terbium(III) was sensitized when excited in the presence of sparfloxacin (SPFX) in the aqueous solution because a Tb(III)-SPFX complex was formed. The sensitized fluorescence was further enhanced when this system was exposed to 365 nm ultraviolet light. By the spectral properties and contrast experiments, it is proved that irradiation makes this system undergo photochemical reactions and a new terbium complex which is more favorable to the intramolecular energy transfer is formed. The mechanism of photochemical fluorescence enhancement of the Tb(III)-SPFX system is discussed and a new sensitive and selective photochemical fluorimetry for the determination of SPFX is established. Under the optimum conditions, the linear range is 1.0-50 x 10(-7) M for SPFX, the detection limit is 3.0 x 10(-9) M and the R.S.D. for 5.0 x 10(-7) M SPFX is 1.3% (n = 9). Without any pretreatment the recovery of SPFX in human urine was determined with satisfaction. (C) 1999 Elsevier Science B.V. All rights reserved.

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The UV-visible absorption and fluorescence spectra of a soluble polyimide, YS-30, in several organic solvents were measured over a wide range of concentration. The experimental results show that there exist both intramolecular and intermolecular electron donor acceptor interactions for YS-30 molecules. The fluorescence behavior of YS-30 in N,N-dimethylacetamide and in chloroform solutions is similar in general, except that its ground-state intermolecular charge transfer emission is more obvious in N,N-dimethylacetamide solution. This difference is attributed to the greater extent of disruption of the chain packing by solvent or/and the more efficient radiationless energy dissipation process from the excited state complexes to chloroform. The intensity ratio of intermolecular charge transfer emission to intramolecular charge transfer emission is used to characterize the state of aggregation of YS-30 molecules in solutions. The plot of this ratio versus concentration indicates the existence of two critical concentrations. It is also found from the same plot that the decrease of coil size is very pronounced during the initial stage of shrinkage.

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Lomefloxacin (LMFX) and terbium ion can form a complex and the sensitized fluorescence of the terbium ion can be observed. It was found that the sensitized fluorescence intensity can be notably enhanced when the terbium complex is exposed to 365 nm ultraviolet light. By the fluorescence spectra, phosphorescence spectra, fluorescence quantum yield and fluorescence lifetime of the system, it was proved that irradiation of the complex made it undergo a photochemical reaction and a new terbium complex which is more favorable to intramolecular energy transfer was formed. This is why the sensitized fluorescence enhancement can be observed. (C) 1999 Elsevier Science S.A. All rights reserved.