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DOI: 10.3791/57592-v
Huang Chunhui1,2, Xu Dilin1, Zhang Ke1, Shentu Jieyi2, Yan Sicheng1, Wu Dapeng3, Wang Qinwen1, Cui Wei1,2
1Ningbo Key Laboratory of Behavioral Neuroscience, Zhejiang Provincial Key Laboratory of Pathophysiology, School of Medicine,Ningbo University, 2Li Dak Sum Yip Yio Chin Kenneth Li Marine Biopharmaceutical Research Center,Ningbo University, 3School of Materials Science and Chemical Engineering,Ningbo University
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This protocol describes the preparation of Aβ oligomers from a synthetic peptide in vitro and the evaluation of their relative amounts using dot blotting analysis. This method is simple, reliable, and reproducible, making it essential for neuropharmacological research.
Este protocolo describe cómo preparar oligómeros de Aß de un péptido sintético en vitro y evaluar cantidades relativas de oligómero Aβ mediante un punto de análisis el borrar.
Este método puede ayudar a responder preguntas clave en el campo neurofarmacológico. La principal ventaja de esta técnica es que es sencilla, fiable y reproducible para formar oligómero beta-amiloide. Mostramos que la demostración de este método es fundamental ya que los pasos para preparar el oligómero beta son difíciles de aprender.
Primero, ajuste la concentración de la solución de monómero A-beta a 0,25 miligramos por mililitro agregando 900 microlitros de agua bidestilada al tubo. Luego continúe evaporando la solución de monómero con gas nitrógeno de alta pureza hasta que el volumen alcance aproximadamente 850 microlitros con una concentración de aproximadamente 0,29 miligramos por mililitro. La presencia de HR5P afecta a la formación de oligómero A-beta.
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