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  A new weakly basic amino-reactive fluorescent label for use in isoelectric focusing and chip electrophoresis

Schulze, P., Link, M., Schulze, M., Thuermann, S., Wolfbeis, O., & Belder, D. (2010). A new weakly basic amino-reactive fluorescent label for use in isoelectric focusing and chip electrophoresis. Electrophoresis, 31(16), 2749-2753. doi:10.1002/elps.201000007.

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 Urheber:
Schulze, P.1, 2, Autor           
Link, M.3, Autor
Schulze, M.1, Autor
Thuermann, S.1, Autor
Wolfbeis, O.S.3, Autor
Belder, D.1, 4, Autor           
Affiliations:
1Institute of Analytical Chemistry, University of Leipzig, Leipzig, Germany, ou_persistent22              
2Service Department Schulze (GC, HPLC), Max-Planck-Institut für Kohlenforschung, Max Planck Society, ou_1445630              
3Institute of Analytical Chemistry, Chemo- and Biosensors, University of Regensburg, Regensburg, Germany, ou_persistent22              
4Research Group Belder, Max-Planck-Institut für Kohlenforschung, Max Planck Society, ou_1445595              

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Schlagwörter: CIEF; Fluorescence marker; MCE; Luminescence detection; Protein labeling
 Zusammenfassung: In this study, we present a novel amino-reactive fluorescence marker (referred to as UR-431), which is well suited for electrophoretic techniques. A main feature of this marker is its weakly basic behavior when conjugated to analytes. Labeled primary amines exhibit a positive net charge and accordingly a cathodic mobility below a pH of 2.4. The label features a pH-independent fluorescence emission and is thus very interesting for electrophoretic applications such as IEF. The absorption maximum of this yellow daylight chromophore is at 431 nm, whereas fluorescence emission peaks at 537 nm (quantum yield≈0.1). The label was successfully conjugated to amines, peptides and proteins and separated via CE and MCE. The on-chip detection limit of labeled lysine using a mercury-lamp-based fluorescence microscope was determined as 12 nM. An important feature of the new label is that it effects only a subtle change of the pI of proteins compared with common anionic labels, e.g. FITC. pI values of proteins were investigated by comparing native proteins and labeled proteins in CIEF. UR-431 was also applied to sensitive detection of amines and peptides in MCE.

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Sprache(n): eng - English
 Datum: 2010-06-192010-08
 Publikationsstatus: Erschienen
 Seiten: -
 Ort, Verlag, Ausgabe: -
 Inhaltsverzeichnis: -
 Art der Begutachtung: Expertenbegutachtung
 Identifikatoren: DOI: 10.1002/elps.201000007
 Art des Abschluß: -

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Titel: Electrophoresis
Genre der Quelle: Zeitschrift
 Urheber:
Affiliations:
Ort, Verlag, Ausgabe: Weinheim, Germany : Verlag Chemie,
Seiten: - Band / Heft: 31 (16) Artikelnummer: - Start- / Endseite: 2749 - 2753 Identifikator: ISSN: 0173-0835
CoNE: https://pure.mpg.de/cone/journals/resource/954925486765