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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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Published in: | Analytical chemistry (Washington) 2018-01, Vol.90 (2), p.1059-1062 |
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container_title | Analytical chemistry (Washington) |
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creator | Nobori, Takanobu Tosaka, Kenta Kawamura, Akira Joichi, Taisei Kamino, Kenta Kishimura, Akihiro Baba, Eishi Mori, Takeshi Katayama, Yoshiki |
description | 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. |
doi_str_mv | 10.1021/acs.analchem.7b03893 |
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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.</description><identifier>ISSN: 0003-2700</identifier><identifier>EISSN: 1520-6882</identifier><identifier>DOI: 10.1021/acs.analchem.7b03893</identifier><identifier>PMID: 29260552</identifier><language>eng</language><publisher>United States: American Chemical Society</publisher><subject>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</subject><ispartof>Analytical chemistry (Washington), 2018-01, Vol.90 (2), p.1059-1062</ispartof><rights>Copyright © 2017 American Chemical Society</rights><rights>Copyright American Chemical Society Jan 16, 2018</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-a525t-c4a2de2eb5b6d193e48e538e2a46ad70af34be9107ca2415a8e4033d435240b3</citedby><cites>FETCH-LOGICAL-a525t-c4a2de2eb5b6d193e48e538e2a46ad70af34be9107ca2415a8e4033d435240b3</cites><orcidid>0000-0002-1821-5427 ; 0000-0002-0503-1418</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/29260552$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Nobori, Takanobu</creatorcontrib><creatorcontrib>Tosaka, Kenta</creatorcontrib><creatorcontrib>Kawamura, Akira</creatorcontrib><creatorcontrib>Joichi, Taisei</creatorcontrib><creatorcontrib>Kamino, Kenta</creatorcontrib><creatorcontrib>Kishimura, Akihiro</creatorcontrib><creatorcontrib>Baba, Eishi</creatorcontrib><creatorcontrib>Mori, Takeshi</creatorcontrib><creatorcontrib>Katayama, Yoshiki</creatorcontrib><title>Alkaline Phosphatase-Catalyzed Amplification of a Fluorescence Signal for Flow Cytometry</title><title>Analytical chemistry (Washington)</title><addtitle>Anal. 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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 |
title | Alkaline Phosphatase-Catalyzed Amplification of a Fluorescence Signal for Flow Cytometry |
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