Alkaline Phosphatase-Catalyzed Amplification of a Fluorescence Signal for Flow Cytometry

Despite the expanding use of flow cytometry, its detection limit is not satisfactory for many antigen proteins with low copy numbers. Herein, we describe an alkaline phosphatase (AP)-based technique to amplify the fluorescence signal for cell staining applications. We designed a fluorescent substrat...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2018-01, Vol.90 (2), p.1059-1062
Main Authors: Nobori, Takanobu, Tosaka, Kenta, Kawamura, Akira, Joichi, Taisei, Kamino, Kenta, Kishimura, Akihiro, Baba, Eishi, Mori, Takeshi, Katayama, Yoshiki
Format: Article
Language:English
Subjects:
A549 Cells
Alkaline phosphatase
Alkaline Phosphatase - metabolism
Amplification
Antigens
Antigens - analysis
Cell Membrane Permeability
Cells
Chemistry
Cytometry
Dephosphorylation
Flow cytometry
Flow Cytometry - methods
Fluorescence
Fluorescent Antibody Technique - methods
Fluorescent Dyes - chemistry
Fluorescent Dyes - metabolism
Humans
Immunoconjugates - chemistry
Immunoconjugates - metabolism
K562 Cells
Membrane permeability
Phosphatase
Phosphorylation
Proteins
Substrates
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Summary:Despite the expanding use of flow cytometry, its detection limit is not satisfactory for many antigen proteins with low copy numbers. Herein, we describe an alkaline phosphatase (AP)-based technique to amplify the fluorescence signal for cell staining applications. We designed a fluorescent substrate that acquires membrane permeability upon dephosphorylation by AP. By using the substrate, the fluorescence signal of cells in flow cytometry could be successfully amplified to give a much stronger signal than the cells labeled using a conventional fluorophore-modified antibody.
ISSN:0003-2700
1520-6882
DOI:10.1021/acs.analchem.7b03893