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GE Healthcare Bio-Sciences AB

2 ARTICLES PUBLISHED IN JoVE

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Biology

Selective Labelling of Cell-surface Proteins using CyDye DIGE Fluor Minimal Dyes
Asa Hagner-McWhirter 1, Maria Winkvist 1, Stephanie Bourin 1, Rita Marouga 1
1Research and Development, GE Healthcare Bio-Sciences AB

A simple and specific method was demonstrated for fluorescent labeling and enhanced detection of cell surface proteins without a fractionation step. Differential abundance in cell surface proteins was analyzed using two-dimensional (2-D) electrophoresis and Ettan™ DIGE technology.

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Biology

The Importance of Correct Protein Concentration for Kinetics and Affinity Determination in Structure-function Analysis
Ewa Pol 1
1GE Healthcare Bio-Sciences AB

We apply label-free protein interaction analysis using Biacore X100 for structure-function analysis of the binding of several cystatin B mutants to papain through kinetic characterization. Calibration-free concentration analysis (CFCA) measures the concentration of protein with retained binding activity without the need for a standard curve. We show that confirmation of concentrations using CFCA increases the reliability of the kinetic analysis and that kinetic constants can reliably be determined even if the activity of a recombinant protein is reduced.

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