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

Schulze, Philipp and Link, Martin and Schulze, Marcel and Thuermann, Sebastian and Wolfbeis, Otto S. and Belder, Detlev (2010) A new weakly basic amino-reactive fluorescent label for use in isoelectric focusing and chip electrophoresis. ELECTROPHORESIS, 31 (16). pp. 2749-2753. ISSN 0173-0835, 1522-2683

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Abstract

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 approximate to 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.

Item Type: Article
Uncontrolled Keywords: SINGLE-MOLECULE DETECTION; CAPILLARY-ELECTROPHORESIS; MICROCHIP ELECTROPHORESIS; MICROFLUIDIC DEVICES; CHAMELEON LABELS; PROTEINS; DYES; SEPARATIONS; DNA; REAGENTS; CIEF; Fluorescence marker; MCE; Luminescence detection; Protein labeling
Subjects: 500 Science > 540 Chemistry & allied sciences
Divisions: Chemistry and Pharmacy > Institut für Analytische Chemie, Chemo- und Biosensorik > Chemo- und Biosensorik (Prof. Antje J. Bäumner, formerly Prof. Wolfbeis)
Depositing User: Dr. Gernot Deinzer
Date Deposited: 20 Jul 2020 08:27
Last Modified: 20 Jul 2020 08:27
URI: https://pred.uni-regensburg.de/id/eprint/24371

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