Comparative study of different fluorescent dyes for the detection of proteins on membranes using the peroxyoxalate chemiluminescent reaction

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• 作者：Salerno ; Doris ; Daban ; Joan-Ramon
• 关键词：Peroxyoxalate chemiluminescence ; Fluorescent dyes ; Proteins
• 刊名：Journal of Chromatography B, Analytical Technologies in the Biomedical and Life Sciences
• 出版年：2003
• 出版时间：August 5, 2003
• 年：2003
• 卷：793
• 期：1
• 页码：75-81
• 全文大小：454 K

文摘

We have previously shown that the bis(2,4,6-trichlorophenyl)oxalate (TCPO)–H₂O₂ chemiluminescent reaction in acetone can be used for the detection of proteins labeled with the fluorescent reagent 2-methoxy-2,4-diphenyl-3(2H)-furanone (MDPF) on polyvinylidene difluoride (PVDF) membranes. To improve this method, in this work we have designed and constructed a cell that allows us to perform this chemiluminescent reaction on PVDF membranes with a homogeneous distribution of the reagents. Using this cell we have examined the analytical properties of several recently developed fluorescent protein dyes chemically different from MDPF. We have found that the metal chelate dye SYPRO Ruby can also be excited by the high-energy intermediate produced in the TCPO–H₂O₂ reaction.