Abstract
This protocol measures the protein-peptide interaction by surface plasmon resonance (SPR) using Biacore X100 (GE Healthcare). The Biacore system can monitor the direct interaction between biomolecules. There are several methods of immobilizing a ligand to the sensor chip. The optimal immobilization method for each experiment needs to be selected. In this protocol, we employed amine coupling to immobilize the protein to the carboxyl-type sensor chip. The procedure generally follows the “Instrument Handbook” of Biacore X100.
Keywords: Signal, Ligand, Histidine kinase, Sensor
Materials and Reagents
Equipment
Procedure
Recipes
The buffers used for immobilization and binding must be optimized for every ligand and analyte. The following recipe worked with our protein and peptide samples.
Acknowledgments
This protocol was adapted from Eguchi et al. (2012), and generally follows the “Instrument Handbook” of Biacore X100 (GE Healthcare). This work was supported by a Grant-in-Aid for Scientific Research (A, 20248012) from the Japan Society for the Promotion of Science (JSPS), the Research and Development Program for New Bio-Industry Initiatives (2006–2010) of Bio-Oriented Technology Research Advancement Institution (BRAIN), Japan, MEXT-Supported Program for the Strategic Research Foundation at Private Universities, 2011-2015 (S1101035), Sasakawa Scientific Research Grant from The Japan Science Society, and the Institute for Fermentation, Osaka.
References
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