Single molecule FRET detection in CdSe-QD donor and Cy5-labeled molecular chaperone acceptor complex by imaging microscopy

Tani, Tadaaki; Oda, Masahiro; Sakai, Hiroshi; Araki, Daisuke; Itoh, Yoshinori; Ohtaki, Akashi; Yohda, Masafumi

doi:10.1016/j.jlumin.2010.09.019

Cited by 3 publications

(2 citation statements)

References 14 publications

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“…QDs and cyanine dye are among the most efficient FRET pairs used over the past decade. 15,25,26 However, most of these FRET systems have generally applied to structural study of larger biomolecules ( i.e. proteins and DNA) and not small glycans.…”

Section: Discussionmentioning

confidence: 99%

“…In addition, the narrow emission spectrum of QD SA resulted in negligible crosstalk between the QD SA and Cy5 fluorophores. QDs and cyanine dye are among the most efficient FRET pairs used over the past decade. ,, However, most of these FRET systems have generally been applied to structural study of larger biomolecules (i.e., proteins and DNA) and not small glycans. Moreover, FRET was mostly detected by imaging and microscopic techniques, which is relatively difficult to apply for quantitative analysis.…”

Section: Discussionmentioning

confidence: 99%

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Ultrasensitive Detection and Quantification of Acidic Disaccharides Using Capillary Electrophoresis and Quantum Dot-Based Fluorescence Resonance Energy Transfer

Chang¹,

Cai²,

Li³

et al. 2013

Anal. Chem.

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Rapid and highly sensitive detection of the carbohydrate components of glycoconjugates is critical for advancing glycobiology. Fluorescence (or Forster) resonance energy transfer (FRET) is commonly used in detection of DNA, in protein structural biology, and in protease assays, but is less frequently applied to glycan analysis due to difficulties in inserting two fluorescent tags into small glycan structures. We report an ultrasensitive method for the detection and quantification of a chondroitin sulfate disaccharide based on FRET, involving a CdSe-ZnS core-shell nanocrystal quantum dot (QD)-streptavidin conjugate donor and a Cy5 acceptor. The disaccharide was doubly labeled with biotin and Cy5. QDs then served to concentrate the target disaccharide, enhancing the overall energy transfer efficiency, with unlinked QDs and Cy5-hydrazide producing nearly zero background signal in capillary electrophoresis using laser-induced fluorescence detection with two different band-pass filters. This method is generally applicable to the ultrasensitive analysis of acidic glycans and offers promise for the high-throughput disaccharide analysis of glycosaminoglycans.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%