title: Control of the fluorescence of dye-antibody conjugates by (2-hydroxypropyl)-β-cyclodextrin in fluorescence microscopy and flow cytometry.
abstract: When proteins are conjugated to fluorescent organic dyes, fluorescence emission of the dye molecules is usually decreased, sometimes up to 50-70%. This quenching phenomenon has been acknowledged for decades, but as yet, there are no simple, practical methods to control the fluorescence of dyes conjugated to proteins, especially for dyes conjugated to immunoglobulins. Here, we report that the addition of (2-hydroxypropyl)-β-cyclodextrin (HPβCD) to dye-antibody conjugates can increase fluorescence up to 2.5-fold in cell imaging and flow analysis. This method may be an effective way to increase the sensitivity of detection of fluorescent organic labels used in immunology, histochemistry, and cell biology.
authors.affiliation: BD Biosciences, 2350 Qume Drive, San Jose, California 95131, United States. oleg_guryev@bd.com
authors.forename: Oleg
authors.initials: O
authors.lastname: Guryev
authors.forename: Barny
authors.initials: B
authors.lastname: Abrams
authors.forename: Chip
authors.initials: C
authors.lastname: Lomas
authors.forename: Qudrat
authors.initials: Q
authors.lastname: Nasraty
authors.forename: Emily
authors.initials: E
authors.lastname: Park
authors.forename: Tim
authors.initials: T
authors.lastname: Dubrovsky
mesh_terms: D000073738:2-Hydroxypropyl-beta-cyclodextrin; D000906:Antibodies / Q000737:chemistry / Q000276:immunology; D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D007136:Immunoglobulins / Q000737:chemistry; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000276:immunology; D008856:Microscopy, Fluorescence / Q000379:methods*; D047392:beta-Cyclodextrins / Q000737:chemistry*
chemical_list: D000906:Antibodies; D005456:Fluorescent Dyes; D007136:Immunoglobulins; D047392:beta-Cyclodextrins; D000073738:2-Hydroxypropyl-beta-cyclodextrin