Europium(III) cryptate: a fluorescent label for the detection of DNA hybrids on solid support.
We report here a new detection method for DNA hybrids on dot blots. The process utilizes DNA or oligonucleotide probes labeled with biotin, followed by recognition with a conjugate of streptavidin and europium cryptate, a time-resolved fluorescent label. Unlike the other lanthanide chelates, this label is an organic molecule embedding a europium ion into an intramolecular cavity. This structure has a better stability in diluted assay media, a good sensitivity even on solid support, and an elevated fluorescence lifetime which allows elimination of most of the background generated by other species present in the assay medium. This procedure is quantitative and detects down to 2 amol of a model DNA, which is similar to other nonisotopic (especially colorimetric) methods. The main advantages of this method are easy automation, quantitation, and rapidity of measurements.
Prat
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O
CIS Biointernational, Laboratoire des Produits pour Analyses Médicales, Bagnols Sur Ceze, France.
Lopez
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E
Mathis
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G
D001426:Bacterial Proteins / Q000032:analysis; D001710:Biotin / Q000032:analysis; D001643:Bridged Bicyclo Compounds / Q000737:chemistry*; D015342:DNA Probes / Q000032:analysis*; D005063:Europium / Q000737:chemistry*; D005456:Fluorescent Dyes; D009693:Nucleic Acid Hybridization; D009942:Organometallic Compounds / Q000737:chemistry*; D019809:Streptavidin
D001426:Bacterial Proteins; D001643:Bridged Bicyclo Compounds; D015342:DNA Probes; D005456:Fluorescent Dyes; D009942:Organometallic Compounds; C070734:europium(III) trisbipyridine cryptate; D005063:Europium; D001710:Biotin; D019809:Streptavidin