The Rab family of small GTPases are key regulators of membrane trafficking in eukaryotic cells. Rab11, one member of this family, plays a role in regulating various cellular functions such as plasma membrane recycling, phagocytosis, and cytokinesis. A family of Rab11-binding proteins has been identified and termed the Rab11 family interacting proteins or Rab11-FIPs. Rab11-FIP3, a member of this Rab11-binding protein family, in addition to interacting with Rab11, is also capable of interaction with members of the ADP-Ribosylation Factor (ARF) GTPase family. Here we describe the purification of Rab11-FIP3 and report its biological properties in eukaryotic cells as visualized by immunofluorescence microscopy.