Begin by immobilizing the purified msRBP4 protein on the sensor chip. To do so, go to the Run menu, choose the wizard, and proceed with the immobilization option. Next, dilute the ligand in immobilization buffer to a concentration of 10 micrograms per milliliter, and aliquot the coupling reagents into vials.
Now, open the Immobilization Setup wizard window. Select the CM5 chip and immobilize flow cell 2 options. Then set the target level to 1200 response units for an R maximum of 30 response units in the kinetic study.
Choose ethanolamine as the wash solution. Enter the name RBP4, and click Next. In the Rack Position dialogue box, click Eject Rack.
Remove the rack. Place the vials containing the specified volumes of each reagent. And then insert the rack with the sample vials.
After that, click OK and Next. Finally, in the Prepare Run Protocol dialogue, click Start and then Save.
