RESUMO
Effects of various factors on synthesis and fluorescent properties of CdSe quantum dots were studied. It was shown that variation of pH, stabilizer and concentration of precursors brings to obtaining of quantum dots with various fluorescent properties. The nanoparticles prepared were conjugated with rabbit antibodies to C-Reactive protein and C-Reactive protein for competitive immunoassay for determination of CRP. It was shown that interaction of these dots as a result of antigen-antibody reaction brings to resonance energy transfer and these changes in fluorescence spectra correlate with concentration of CRP. This approach permits to determine CRP in range between 4-100 ng.
Assuntos
Proteína C-Reativa/análise , Compostos de Cádmio/química , Fluorescência , Imunoensaio , Pontos Quânticos , Compostos de Selênio/química , Água/química , Humanos , SolubilidadeRESUMO
Affinity purification of annexin V from human placenta on column with appropriate monospecific antibodies is developed. The procedure permits purification of the protein to a highly purified state by a two stage procedure. The yield of the protein is about 5 mg per 100 g of wet tissue. Because of high homologies between various annexins, it was supposed that this procedure can be also applied for purification of other annexins from other tissues.
Assuntos
Anexina A5/isolamento & purificação , Cromatografia de Afinidade/métodos , Placenta/metabolismo , HumanosRESUMO
Lectin binding assay was applied to polyacrylamide slices to study the effects of lysozyme on carbohydrate profile of plasma membranes from liver and heart tissues. The investigations were carried out on polyacrylamide slices immobilized with wheat germ agglutinin or Concanavalin A. It was shown that lysozyme changes the carbohydrate profile of plasma membranes of liver and heart cells.
Assuntos
Membrana Celular/efeitos dos fármacos , Coração/efeitos dos fármacos , Hepatócitos/efeitos dos fármacos , Lectinas/metabolismo , Muramidase/farmacologia , Resinas Acrílicas/química , Animais , Carboidratos/análise , Membrana Celular/química , Sobrevivência Celular/efeitos dos fármacos , Técnicas de Laboratório Clínico , Concanavalina A/química , Concanavalina A/metabolismo , Hepatócitos/ultraestrutura , Miocárdio/citologia , Miocárdio/ultraestrutura , Ligação Proteica , Ratos , Extratos de Tecidos/química , Aglutininas do Germe de Trigo/químicaRESUMO
Polystyrene latex beads were polymerized at two different pH (7.3 and 9.5) and their ability to create latex agglutination systems were studied. Sensitization of these latexes by DNA, antibodies to C-reactive protein, myoglobin, and human IgG. was conducted. Sensitivity and specificity of these systems were compared. Practical recommendations are given for preparation of such systems.
Assuntos
Testes de Fixação do Látex/normas , Poliestirenos/síntese química , Animais , Anticorpos , Antígenos , Proteína C-Reativa/imunologia , Bovinos , DNA , Humanos , Interações Hidrofóbicas e Hidrofílicas , Imunoglobulina G/imunologia , Microesferas , Mioglobina/imunologia , Poliestirenos/normas , Ligação Proteica , Coelhos , Sensibilidade e EspecificidadeRESUMO
A simplified procedure for purification of annexin V from human placenta was developed. At first, the protein was separated from other proteins in membrane bound form in the presence of Ca2+, then was extracted with EDTA and purified by affinity chromatography on PAAG-immobilized phosphatidylserine. The purified protein gave a single band with a molecular weight of 35,000 in SDS-PAGE.