Phosphoprotein staining for sodium dodecyl sulfate-polyacrylamide gel electrophoresis using fluorescent reagent morin hydrate

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• 作者：Xu Wang^a ; Sun-Young Hwang^a ; Wei-Tao Cong^b ; Li-Tai Jin^b ; Jung-Kap Choi^a ; ^{jkchoi@chonnam.ac.kr}
• 关键词：Phosphoprotein stain ; Fluorescent ; Morin hydrate ; Aluminum ; Pro-Q Diamond ; SDS&ndash ; PAGE
• 刊名：Analytical Biochemistry
• 出版年：2013
• 出版时间：1 April, 2013
• 年：2013
• 卷：435
• 期：1
• 页码：19-26
• 全文大小：1023 K

文摘

A fluorescence-based stain with 3,5,7,2¡ä,4¡ä-pentahydroxyflavone (morin hydrate, MH) was designed to stain phosphoproteins in one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Al³⁺ was applied as a ¡°fixed bridge,¡± providing an efficient energy transfer channel between phosphoprotein and MH, to produce a strong fluorescent complex for the determination of phosphoprotein. As little as 62.5 ng of ¦Á-casein (7 or 8 phosphates) and ¦Â-casein (5 phosphates), 125 ng of ovalbumin (2 phosphates), and ¦Ê-casein (1 phosphate) could be visualized with a wide linear dynamic range. In comparison with conventional methods, MH stain is a time-saving method that takes just 90 min. It also has good compatibility with routine protein stainings such as Coomassie Brilliant Blue R (CBBR) and SYPRO Ruby for total protein analysis.