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Purification of human endothelial overexpressed lipopolysaccharide-associated factor 1 protein / 中华烧伤杂志
Chinese Journal of Burns ; (6): 367-369, 2005.
Artigo em Chinês | WPRIM | ID: wpr-312540
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the feasibility of obtaining of a highly pure protein of human endothelial overexpressed lipopolysaccharide-associated factor 1 (EOLA1) with metal chelation chromatography.</p><p><b>METHODS</b>Inclusion bodies of the E. coli transformed with EOLA1 gene were extracted and washed with BugBuster Protein Extraction Reagent. The primary purified products were purified by His. Bind Resin Chromatography under denaturing condition and dialyzed for renaturation, and then were analyzed with SDS-PAGE, Western blotting and peptide mass fingerprinting (PMF).</p><p><b>RESULTS</b>EOLA1 was mainly expressed in E. coli as insoluble inclusion bodies. The protein content in the primary extracted inclusion bodies accounted for over 75%, and it accounted for more than 90% after chromatography and renaturation. It was indicated by PMF that the targeted protein peptide overlaid many of designed protein peptide.</p><p><b>CONCLUSION</b>The method of EOLA1 protein purification and renaturation was convenient and efficient, and by this method sufficient amount of highly pure EOLA1 protein could be obtained for the preparation of EOLA1 monoclonal antibody and for the study of its gene function.</p>
Assuntos
Texto completo: DisponíveL Índice: WPRIM (Pacífico Ocidental) Assunto principal: Immunoblotting / Cromatografia de Afinidade / Células Endoteliais / Eletroforese em Gel de Poliacrilamida / Genética / Proteínas de Membrana / Metabolismo / Anticorpos Monoclonais Limite: Humanos Idioma: Chinês Revista: Chinese Journal of Burns Ano de publicação: 2005 Tipo de documento: Artigo

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Texto completo: DisponíveL Índice: WPRIM (Pacífico Ocidental) Assunto principal: Immunoblotting / Cromatografia de Afinidade / Células Endoteliais / Eletroforese em Gel de Poliacrilamida / Genética / Proteínas de Membrana / Metabolismo / Anticorpos Monoclonais Limite: Humanos Idioma: Chinês Revista: Chinese Journal of Burns Ano de publicação: 2005 Tipo de documento: Artigo