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2 articles published in JoVE
The Importance of Correct Protein Concentration for Kinetics and Affinity Determination in Structure-function Analysis Ewa Pol1 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.
Selective Labelling of Cell-surface Proteins using CyDye DIGE Fluor Minimal Dyes Asa Hagner-McWhirter1, Maria Winkvist1, Stephanie Bourin1, Rita Marouga1 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.