Munich University of Applied Sciences

2 ARTICLES PUBLISHED IN JoVE

Biochemistry

Covalent Immobilization of Proteins for the Single Molecule Force Spectroscopy

Tanja D. Becke^1,2,3, Stefan Ness², Stefanie Sudhop^1,3, Hermann E. Gaub³, Markus Hilleringmann², Arndt F. Schilling^*4, Hauke Clausen-Schaumann^*1,3

¹Center for Applied Tissue Engineering and Regenerative Medicine, Munich University of Applied Sciences, ²FG Protein Biochemistry & Cellular Microbiology, Munich University of Applied Sciences, ³Center for Nano Science, Ludwig-Maximilians-Universität München, ⁴Klinik für Unfallchirurgie, Orthopädie und Plastische Chirurgie, University Medical Center Göttingen

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.

Bioengineering

Extracting the Young's Modulus of Native Murine Pulmonary Basement Membranes from Atomic Force Microscopy derived Force Maps

Bastian Hartmann^1,2, Monica Nicolau^3,4, Raphael Reuten^*3,4, Hauke Clausen-Schaumann^*1,2

¹Munich University of Applied Sciences, ²Center for Applied Tissue Engineering and Regenerative Medicine - CANTER, Munich University of Applied Sciences, ³Experimental and Clinical Pharmacology and Toxicology, Medical Faculty, University of Freiburg, ⁴Department of Obstetrics and Gynecology, Medical Center, University of Freiburg

This protocol visualizes how to prepare cryosections of murine lung tissue, perform atomic force microscopy force map experiments, and analyze the data to determine Young's modulus of the native murine pulmonary basement membrane.