Related Experiment Video
Updated: Jul 3, 2026
Double Labeling Immunofluorescence using Antibodies from the Same Species to Study Host-Pathogen Interactions
Published on: July 10, 2021
Effect of fluorochrome location in protein A on sensing efficiency of IgG
1Department of Chemical and Biochemical Engineering, University of Maryland Baltimore County Campus, Baltimore, Maryland 21228, USA.
Fragment B of protein A conjugated with fluorescein at various lysine residues is prepared and separated by using a DEAE column in anion-exchange chromatography. The binding of IgG Fc to fragment B contributes to an additional positive electric potential around fragment B. The change in the local electrostatic environment and pH can then be specifically monitored by measuring the fluorescence intensity of fluorescein conjugated with fragment B before and after the introduction of IgG. The studies for the quantitative dependence of fluorescein location on the effectiveness of fluorescein for sensing the protein A-IgG reaction are presented and discussed.
Fragment B of protein A conjugated with fluorescein at various lysine residues is prepared and separated by using a DEAE column in anion-exchange chromatography. The binding of IgG Fc to fragment B contributes to an additional positive electric potential around fragment B. The change in the local electrostatic environment and pH can then be specifically monitored by measuring the fluorescence intensity of fluorescein conjugated with fragment B before and after the introduction of IgG. The studies for the quantitative dependence of fluorescein location on the effectiveness of fluorescein for sensing the protein A-IgG reaction are presented and discussed.
