Biophysics Lab Course, Institute of Biophysics
Idea: In this experiment we determine the binding of a ligand to a receptor using spectroscopic means.
The amino acid Trp-214 within HSA (human serum albumin) acts as a FRET donor and the ligand 3-hydroxyflavone acts as the acceptor.
Acceptor ligand is successively added and the decreasing donor fluorescence and increasing acceptor fluorescence due to FRET-complexes is recorded.
The binding affinity is investigated from both the donor-quenching and analysis of the average FRET-efficiency in a binding isotherm. The latter also gives information about the D-A distance.
Experiments: Gene expression | Bioinformatics | Protein labelling | Protein crystallization | Protein folding | Stopped-flow kinetics | Single-molecule fluorescence | Fluorescence lifetime | Ligand binding | Fluorescence correlation spectroscopy (FCS) | Lipid monolayers | Ion channels | Live cell imaging | Superresolution microscopy | Optical tweezers | AFM
