This protocol describes the covalent immobilization of proteins with a heterobifunctional silane coupling agent to silicon-oxide surfaces designed for the atomic force microscopy based single molecule force spectroscopy which is exemplified by the interaction of RrgA (pilus-1 tip adhesin of S. pneumoniae) with fibronectin.
Becke, T. D., Ness, S., Sudhop, S., Gaub, H. E., Hilleringmann, M., Schilling, A. F., Clausen-Schaumann, H. Covalent Immobilization of Proteins for the Single Molecule Force Spectroscopy. J. Vis. Exp. (138), e58167, doi:10.3791/58167 (2018).