Surface plasmon resonance (SPR) is an optical sensing method for label-free detection of biomolecular interactions. The method is based on immobilization of a ligand on the sensor surface and reinjection of the analyses onto the surface containing the ligand. After injection, binding is monitored by changes in the refractive index of the medium close to the sensor surface. It is commonly used for detailed and quantitative studies of protein-protein interactions as well as kinetics.
We introduce the basic concepts of SPR and how it can be used to analyze and visualize biomolecular interactions. We provide SPR specific experimental protocols and troubleshooting for drug development, life science research, and other fields.
|Stages||Solutions and Reagents|
|Preparation||Diluting solution, ligand, analyte|
|Immobilization||Running buffer, activation buffer, immobilization buffer, stabilization buffer,|
Express and purify ligands and analytes. Check the purity and stability of the protein. Prepare various materials and buffers for later use.
Select a suitable sensor chip. Perform conditioning and activation of the sensor chip to form reactive groups on the sensor chip surface. Which sensor chip to use depends on the degree of immobilization of the desired ligand and the specific application. Determine the optimal pH of the immobilization buffer and the concentration of the ligand. Inject the ligand protein with the desired injection parameters to achieve immobilization.
Measure the concentration of the analyte. Dilute the analyte using the running buffer. Place the sample in the selected position and insert it into the sample holder. Inject the analyte for binding analysis.
It is possible to select different modes of analysis on the instrument. Select binding analysis and direct binding to study protein binding. Select kinetic analysis and choose mass transfer to run kinetic analysis and binding experiments. Perform data analysis after data is obtained.
Negative binding signals
Regeneration problems in SPR instrument
We will be happy to discuss the details of your anticipated surface plasmon resonance and develop an experimental approach based on your requirements. Please review the SRP service on our website and contact Creative Biolabs.
For research use only. Not intended for any clinical use.