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

We have previously shown that the bis(2,4,6-trichlorophenyl)oxalate (TCPO)–H 2O 2 chemiluminescent reaction in acetone can be used for the detection of proteins labeled with the fluorescent reagent 2-methoxy-2,4-diphenyl-3(2 H)-furanone (MDPF) on polyvinylidene difluoride (PVDF) membranes. To improv...

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Published inJournal of chromatography. B, Analytical technologies in the biomedical and life sciences Vol. 793; no. 1; pp. 75 - 81
Main Authors Salerno, Doris, Daban, Joan-Ramon
Format Journal Article
LanguageEnglish
Published Netherlands Elsevier B.V 05.08.2003
Subjects
Fluorescent Dyes - chemistry
Membranes, Artificial
Oxalates - chemistry
Polyvinyls - chemistry
Proteins
Proteins - analysis
Proteins
Fluorescent dyes
Peroxyoxalate chemiluminescence
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Summary:We have previously shown that the bis(2,4,6-trichlorophenyl)oxalate (TCPO)–H 2O 2 chemiluminescent reaction in acetone can be used for the detection of proteins labeled with the fluorescent reagent 2-methoxy-2,4-diphenyl-3(2 H)-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–H 2O 2 reaction.
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ISSN:1570-0232
1873-376X
DOI:10.1016/S1570-0232(03)00365-9