Abstract
The analytical ultracentrifugation (AUC) is a classic biophysical instrument to analyze protein interactions in solutions. A recently introduced fluorescence detection system (FDS) improves the specificity and sensitivity of the AUC, and offers the potential to analyze the protein interactions in biological fluids. We explore challenges posed by the application of FDS-AUC to the study of protein interactions in E.coli whole cell extracts, using fluorescent proteins (EGFPs) as model system. At experimentally feasible concentrations of cell extracts, we find no discernable effects of hydrodynamic nonideality on the sedimentation of EGFPs. However, at high concentrations E.coli whole cell extracts produce significant signals from auto fluorescence with complicated quenching patterns. Goal of this work is to establish detection limits and develop procedures to improve specificity.
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.