The polarized epithelial cells that comprise the proximal tubule (PT) have a specialized apical endocytic pathway that allows for the high-capacity endocytosis necessary to recover essential nutrients and to maintain a protein-free urine. Impairments in this pathway result in tubular proteinuria, which can worsen to cause more severe kidney disease. Megalin and cubilin, multi-ligand receptors at the apical surface of the epithelial cells, bind proteins in the ultrafiltrate and internalize them via receptor-mediated endocytosis. Ligands are sorted from receptors in endocytic compartments, and the receptors are recycled back to the surface through structures called dense apical tubules (DATs). The molecular identities of the compartments involved in sorting and recycling in PT cells and the kinetics of megalin trafficking through them are unknown. Understanding megalin’s endocytic trafficking itinerary and the key regulatory steps in this pathway is important for discerning the basis of proteinuric diseases and devising therapies. To address this, we identified endocytic compartment markers in a previously developed opossum kidney (OK) cell culture model that recapitulates morphologic and functional features of the PT in vivo. The fraction of total megalin colocalizing with each compartment marker was quantified by Mander’s coefficient from deconvolved confocal images.  Most megalin is localized primarily in Rab11-positive compartments at steady state while a much smaller fraction of megalin is localized in Rab4-positive structures. Surface biotinylation based assays revealed that only a small fraction of total megalin is present at the apical surface at steady state and that megalin is rapidly internalized from the surface. The half-life of surface megalin was also assessed. Our biochemical and quantitative colocalization data was used to construct and refine a kinetic model of megalin trafficking in the PT. These data suggest that recycling is the rate-limiting step in regulating the fraction of total megalin available at the surface. Together, our results provide valuable insight into the spatial organization of the endolysosomal system and the itinerary and regulation of megalin traffic in the PT. Our kinetic model will be used to identify how genetic mutations and other conditions that cause tubular proteinuria disrupt traffic along the endocytic pathway.