UTHSCSA Center for Surface Plasmon Resonance (CSPR)

Initial Consult

Goals and responsibilities

Our goal is to provide the researcher with the highest quality data possible with the obvious benefits to both the researcher and the Center for Surface Plasmon Resonance (CSPR). The researcher is responsible for providing high quality samples for study and may be required to analyze samples by light scattering, spectrophotometry, and/or sedimentation velocity (analytical ultracentrifugation) prior to study with surface plasmon resonance. These technologies are available in the Center for Macromolecular Interations (CMMI).

Worksheet

Please describe the interactions you wish to study using SPR, the questions to which you require answers (equilibrium, kinetics, yes/no), and provide whatever background information is available by others who studied the same or similar interactions. Please email references (pdf copies preferred) to the Technical Director.

Please list the samples you wish to examine with SPR to include names, MW, estimated quantity, and concentration of each.

Initial Consult

Once all the information has been gathered and preliminary experiments to show quality of samples have been performed to the satisfaction of the Director and Technical Director of CSPR, we will meet with the PI and laboratory personnel to discuss an experimental protocol to address their research questions. Initial studies may be performed to assess the feasibility of further studies and will focus on whether or not the molecules show non-specific binding, pH at which to couple ligand, whether binding occurs between the ligand and analyte(s) and in what analyte concentration range, and how to optimally regenerate the surface.

Preparation of samples and buffers

All solutions must be filtered (0.22µ filter) and thoroughly degassed prior to use. Although not the only buffer, the workhorse running buffer is HEPES buffered saline, pH 7.4 (10 mM HEPES, pH 7.4; 150 mM NaCl) containing 3 mM EDTA and 0.005% surfactant P20. This buffer may be prepared or purchased. If prepared, the surfactant is added after filtering and degassing. Analytes must be dialyzed into the chosen running buffer and the dialysis buffer must be used for analyte dilutions and as the running buffer to prevent buffer mismatching. All samples [ligand(s) and analyte(s)] are centrifuged on the day of use at 100,000 rpm for 10 minutes (a table top ultracentrifuge in Dr. Lafer's research laboratory is available to CSPR users with permission of her laboratory personnel).