Effect of Fluorescently Labeling Protein Probes on Kinetics of Protein−Ligand Reactions Export

@article{citeulike:3499876, abstract = {Abstract: We studied the effect of fluorescently labeling proteins on proteinligand reactions. Unlabeled ligands (streptavidin-binding peptides and rabbit immunoglobulin G (IgG) as antigen targets) are immobilized on epoxy-functionalized glass slides. Unlabeled and Cy3-labeled protein probes from the same batch (streptavidin and goat antibodies) subsequently react with the surface-immobilized targets. By monitoring in situ the surface mass density change using an oblique-incidence reflectivity difference scanning microscope (a label-free detector), we measured kon and koff for streptavidinpeptide reactions and antibodyantigen reaction. We found that (1) equilibrium dissociation constants, defined as KD = koff/kon, for streptavidinpeptide reactions increases by a factor of 34 when the solution-phase streptavidin is labeled with Cy3 dye and (2) KD for reactions of solution-phase goat anti-rabbit antibodies with rabbit IgG targets also change significantly when the goat antibodies are labeled with Cy3 dye.}, address = {School of Medicine, University of California at Davis, Sacramento, California 95817}, author = {Sun, Y. S. and Landry, J. P. and Fei, Y. Y. and Zhu, X. D. and Luo, J. T. and Wang, X. B. and Lam, K. S.}, citeulike-article-id = {3499876}, citeulike-linkout-0 = {http://dx.doi.org/10.1021/la802097z}, citeulike-linkout-1 = {http://pubs.acs.org/doi/abs/10.1021/la802097z}, day = {8}, doi = {10.1021/la802097z}, journal = {Langmuir}, keywords = {binding, effect, fluorescent, labeling, spr, sun08pdf}, month = {November}, posted-at = {2008-11-10 02:39:42}, priority = {2}, title = {Effect of Fluorescently Labeling Protein Probes on Kinetics of Protein\&\#x2212;Ligand Reactions}, url = {http://dx.doi.org/10.1021/la802097z}, year = {2008} }

TY - JOUR ID - citeulike:3499876 L3 - citeulike-article-id:3499876 N2 - Abstract: We studied the effect of fluorescently labeling proteins on proteinligand reactions. Unlabeled ligands (streptavidin-binding peptides and rabbit immunoglobulin G (IgG) as antigen targets) are immobilized on epoxy-functionalized glass slides. Unlabeled and Cy3-labeled protein probes from the same batch (streptavidin and goat antibodies) subsequently react with the surface-immobilized targets. By monitoring in situ the surface mass density change using an oblique-incidence reflectivity difference scanning microscope (a label-free detector), we measured kon and koff for streptavidinpeptide reactions and antibodyantigen reaction. We found that (1) equilibrium dissociation constants, defined as KD = koff/kon, for streptavidinpeptide reactions increases by a factor of 34 when the solution-phase streptavidin is labeled with Cy3 dye and (2) KD for reactions of solution-phase goat anti-rabbit antibodies with rabbit IgG targets also change significantly when the goat antibodies are labeled with Cy3 dye. JF - Langmuir AD - School of Medicine, University of California at Davis, Sacramento, California 95817 TI - Effect of Fluorescently Labeling Protein Probes on Kinetics of Protein−Ligand Reactions KW - binding KW - effect KW - fluorescent KW - labeling KW - spr KW - sun08pdf AU - Sun, YS AU - Landry, JP AU - Fei, YY AU - Zhu, XD AU - Luo, JT AU - Wang, XB AU - Lam, KS PY - 2008/11/08/ UR - http://dx.doi.org/10.1021/la802097z ER -