Surface plasmon resonance is particularly important due to the ability of biomolecules of interest to interact with a specific binding partner, and may therefore be more informative than generic measurement techniques. A growing number of robust and reproducible immobilized surfaces chips are available from Biacore, allowing us to analyze various biomolecular interactions. Here we describe a protocol by which the Biacore chips may be reused multiple times to analyze biomolecular interactions without interfering analysis from previously used surfaces or analytes. This procedure will not only help to extend the lifetime of these chips but at the same time render them to be more commercially affordable, especially in a resource-poor setting. The time range for the entire protocol is ∼1 day, including stripping off previously immobilized materials and re-functionalization of gold surface.