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Application of two-color infrared fluorescence for detection of protein phosphorylation / 南方医科大学学报
Journal of Southern Medical University ; (12): 150-153, 2006.
Artigo em Chinês | WPRIM | ID: wpr-234174
ABSTRACT
<p><b>OBJECTIVE</b>To assess the value of two-color infrared fluorescence imaging system in detecting protein phosphorylation in comparison with chemiluminescent detection.</p><p><b>METHOD</b>The lung tissue homogenate of mice treated with lipopolysaccharide (LPS) for different time lengths were prepared to separate the proteins by SDS-polyacrylamide gel electrophoresis followed by transfer of the proteins onto PVDF membranes. The membranes were incubated with the antibodies against total p42/44 MAPK/phospho-p42/44 MAPK, and then with goat anti-mouse or anti-rabbit secondary antibodies conjugated to Alexa Fluor 680, IRDye 800 or horseradish peroxidase. The blotted proteins were detected and quantified using Odyssey infrared imaging system or chemiluminescent detection.</p><p><b>RESULTS</b>LPS treatment rapidly induced p42/44 MAPK phosphorylation, which reached the peak level 1 h after the treatment and resumed the baseline level at 12 h. Consistent results were obtained by the two detection methods, but two-color infrared fluorescence imaging system showed better sensitivity in detecting the target protein, and was easy to manipulate with good efficiency and capable of analyzing two proteins simultaneously.</p><p><b>CONCLUSION</b>Two-color infrared fluorescence system is a powerful system for detecting phosphorylation of proteins.</p>
Assuntos
Texto completo: DisponíveL Índice: WPRIM (Pacífico Ocidental) Assunto principal: Patologia / Fosforilação / Choque Séptico / Química / Lipopolissacarídeos / Western Blotting / Reprodutibilidade dos Testes / Proteína Quinase 1 Ativada por Mitógeno / Proteína Quinase 3 Ativada por Mitógeno / Fluorescência Tipo de estudo: Estudo diagnóstico Limite: Animais Idioma: Chinês Revista: Journal of Southern Medical University Ano de publicação: 2006 Tipo de documento: Artigo

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Texto completo: DisponíveL Índice: WPRIM (Pacífico Ocidental) Assunto principal: Patologia / Fosforilação / Choque Séptico / Química / Lipopolissacarídeos / Western Blotting / Reprodutibilidade dos Testes / Proteína Quinase 1 Ativada por Mitógeno / Proteína Quinase 3 Ativada por Mitógeno / Fluorescência Tipo de estudo: Estudo diagnóstico Limite: Animais Idioma: Chinês Revista: Journal of Southern Medical University Ano de publicação: 2006 Tipo de documento: Artigo