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(2)O(2) 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-H(2)O(2) reaction.
Salerno
Doris
D
Departament de Bioquímica i Biologia Molecular, Facultat de Ciències, Universitat Autònoma de Barcelona, 08193, Bellaterra, Barcelona, Spain.
Daban
Joan-Ramon
JR
D005456:Fluorescent Dyes / Q000737:chemistry*; D008567:Membranes, Artificial; D010070:Oxalates / Q000737:chemistry*; D011145:Polyvinyls / Q000737:chemistry; D011506:Proteins / Q000032:analysis*; D005465:Fluorocarbon Polymers
D005456:Fluorescent Dyes; D008567:Membranes, Artificial; D010070:Oxalates; D011145:Polyvinyls; D011506:Proteins; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers; C044622:peroxyoxalate