246 resultados para Eletroforese
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Pós-graduação em Química - IBILCE
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Pós-graduação em Química - IQ
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Pós-graduação em Química - IQ
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Pós-graduação em Microbiologia - IBILCE
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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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Mesenchymal stem cells (MSCs) are a heterogeneous population of cells that proliferate in vitro as plastic-adherent cells, have fibroblast-like morphology and can differentiate into bone, cartilage and fat cells. Therapeutic potential of MSCs have been studied in experimental models, such as rabbit, in Laboratory of Cell Engineering of Botucatu. However, no specific markers have been reported for expanded rabbit MSCs, which hampers the isolation of pure MSC populations by immunophenotypic characterization. Thus, the objective of this study was to produce monoclonal antibodies (mAbs) to rabbit MSCs. MSCs derived from rabbit bone marrow (BM) were isolated, cultured, expanded ex vivo, and immunized into three BALB/c mices, and spleen cells subsequently harvested were used to generate hibridoma cell lines secreting antibodies against MSCs. Hybridoma cells were screened by flow cytometry and antibody-producing cells were subjected to subsequent rounds of retests. MSC1-160 obtained the best positivity for IgG expression and was cloned by limiting dilutions and micromanipulation. Ascitic fluid from ten best clones was purified by affinity chromatography in Protein A-sepharose CL-4B column and purification control was performed by electrophoresis in agarose gels. The purified IgG were tested against rabbit MSCs, obtaining high positivity by flow Cytometry. In conclusion, we developed 10 mAbs, MSC1-160 A20, A30, A41, A47, A55, A60, A63, A69, A81, and A82, that recognize rabbit MSC cell surface antigens showing potential for immunophenotypic characterization of rabbit MSC cell lines
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Junto ao Laboratório de Fracionamento de Proteínas, desenvolveu atividades aplicando técnicas eletroforéticas (gel de agarose, eletroforese nativa em gel de poliacrilamida) em amostras de músculos de aves de interesse comercial (frangos, perus e avestruzes), tendo como co-orientador o Prof. Paulo Tadeu Figueira da Faculdade de Medicina Veterinária, PUC/PR - Toledo, onde desenvolveu atividades de análise de géis de eletroforese. Aplicou as técnicas de eletroforeses citadas em amostras de músculos e soro sanguíneo (em andamento) de tilápias (Oreochromis niloticus), bem como desenvolveu atividades de análises computacionais (VDS – PHARMACIA) e estatísticas (ASSISTAT) nos dados obtidos, resultando na participação em 7 congressos com apresentação de trabalhos. Cursou a disciplina de Genética, como aluno especial, junto à turma de Licenciatura em Ciências Biológicas, num total de 8 créditos, sendo aprovado com média 7,2
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Canine Pyometra is a uterine disease that occurs in sexually mature bitches, with higher incidence in nulliparous and animals over 4 years and is characterized by presenting an accumulation of pus in the uterine lumen, usually occurring in diestrus. Laboratory tests are important tools for the detection of metabolic abnormalities associated with sepsis and renal function, which are serious consequences of pyometra. In blood the main findings are normochromic non-regenerative anemia, presence of dehydration, and sometimes thrombocytopenia. The WBC count may be normal but most often occurs a neutrophilic leukocytosis with a left shift, monocytosis and the presence of toxic neutrophils. In less than 1 / 3 of the animals the presence of azotemia is present and a density lower than 1035 is detected in the urine of almost 90% of bitches which may be in normal range at the onset of the desease. Urinary protein loss is rare but the protein may be elevated in the reagent strip due to urinary contamination by uterine secretion. The increase of gamma-glutamyltransferase (GGT), alkaline phosphatase (ALP), aspartate aminotransferase (AST) and creatine kinase (CK) may be present, indicating disorders in the liver. Currently, additional laboratory tests are being studied for the diagnosis of pyometra and its prognosis, such as the measurement of C-reactive protein and fibrinogen for monitoring the recovery of the inflammatory process and the urine electrophoresis to characterize the origin of proteinuria in these animals . The aim of this work is to review the literature on the main laboratory tests that aid the diagnosis of Pyometra
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A glicerol quinase é uma proteína tetramérica que cataliza a fosforilação do glicerol a glicerol-3-fosfato, composta por quatro subunidades - sua análise eletroforética fornecerá apenas uma banda se for constatada a sua pureza num gel não desnaturante - e sua reação com o glicerol é dependente de magnésio e de ATP. A eletroforese de proteínas utiliza como suporte um gel de poliacrilamida, que é formado pela polimerização de monômeros de acrilamida ao longo da sua cadeia e ligações cruzadas de cadeias pela inclusão de um co-monômero bifuncional apropriado, usualmente a N,N’ – metileno-bis-acrilamida ou apenas Bis . A eletroforese de proteínas mais comum é a que utiliza SDS para um gel de poliacrilamida desnaturante. O SDS é um sal chamado Dodecil Sulfato de Sódio que tem por característica se ligar a cadeia proteína nos resíduos de aminoácidos apolares, deixando com a carga negativa, sendo assim, toda proteína fica com carga total negativa, migrando para o anodo na eletroforese. As técnicas de purificação utilizadas foram ultrafiltração e precipitação com ácido tricloro acético, etanol gelado e sulfato de amônio. A dupla precipitação com etanol resultou na recuperação de maior quantidade de proteínas. A coloração do gel com prata foi mais sensível do que Comassie blue. O gel de eletroforese mostrou quatro bandas, correspondentes às quatro subunidades da glicerol quinase quando revelados com prata em gel de SDS-PAGE.
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The food industry has been rapidly modernized, and with this the disposal and the consumption of industrialized food has been increasing continually. These products lose their original morphological characteristics, requiring fast and reliable tests that could help to identify the species in question, as most fraudulent behavior in the milk and dairy industry (meat and fish) is carried out where there is partial or total exchange of the original material for other with less value at market. Nowadays there is a lot of techniques that can be used for the identification of animal species, based on muscle protein, or DNA analysis. In the case of protein based analysis, we can mention several types of electrophoresis and immunologic methods, as ELISA. In the case of DNA based methods, we have several assays that use the amplification of DNA fragments, known as PCR, as proof. All these techniques have advantages and disadvantages that can be affected by factors- the sample condition, or the degree of relation between the species in question. Because of this, it’s necessary that a continuous study looking for the improvement of the available techniques, making sure that the confirmation of food authenticity is in place. This is to ensure the true product value, to comply with labeling regulationand and protect the consumer of frauds