For kinetic experiments, choosing the appropriate flow rate can be a crucial factor for determining the correct affinity. Insufficient flow rates can potentially lead to measurement artifacts. During the association phase, a too slow flow rate can be the cause for mass transport limitation effects. During the dissociation phase an inadequate flow rate can result in re-binding effects. Both these effects should be avoided to obtain high quality kinetic data. Thus, the general rule is to work at high flow rates (> 100 µl/min) whenever the experimental circumstances allow.
Category:
Experimental Considerations