Septins are a family of GTP binding proteins implicated in mammalian cell division Most studies examining the role of septins in this process have treated the family as a whole, thus neglecting the possibility that individual members may have diverse functions To address this, we individually depleted each septin family member expressed in He La cells by siRNA and assayed for defects in cell division by immuno fluorescence and time lapse microscopy Depletion of SEPT2, SEPT7, and SEPT11 causes defects in the early stages of cytokinesis, ultimately resulting in binucleation In sharp contrast, SEPT9 is dispensable for the early stages of cell division, but is critical for the final separation of daughter cells Rescue experiments indicate that SEPT9 isoforms containing the N terminal region are sufficient to drive cytokinesis We demonstrate that SEPT9 mediates the localization of the vesicle tethering exocyst complex to the midbody, providing mechanistic insight into the role of SEPT9 during abscission