Hypertension provokes differential trafficking of renal proximal tubule Na⁺/H⁺ exchanger 3 (NHE3) to the base of the apical microvilli and Na⁺-Pi co-transporter 2 (NaPi2) to endosomes. The resultant diuresis and natriuresis are key to blood pressure control. We tested the hypothesis that this differential trafficking of NHE3 vs. NaPi2 was associated with partitioning to distinct membrane domains. In anesthetized rats, arterial pressure was increased (104±2 to 142±4mmHg, 15min) by arterial constriction and urine output increased 23-fold. Renal membranes were fractionated by cold 1% Triton X-100 extraction then centrifugation through OptiPrep flotation gradients. In controls, 84±9% of NHE3 localized to flotillin-enriched lipid raft domains and 69±5% of NaPi2 localized to transferrin receptor-enriched non-rafts. MyosinVI and DPPIV, associated with NHE3 regulation, co-enriched in lipid rafts with NHE3 while NHERF-1 co-enriched in non-rafts with NaPi2. Partitioning was not altered by hypertension. Detergent insoluble (DI) membranes were pelleted after detergent extraction. NHE3 detergent insolubility decreased as it redistributed from body (80±10% DI) to base (75±3%) of the apical microvilli, while NaPi2 partitioned into more insoluble domains as it moved from the microvilli (45±7% DI) to endosomes (82±1%). In conclusion, NHE3 and NaPi2, while both localized to apical microvilli, are segregated into domains: NHE3 to lipid rafts and NaPi2 to non-rafts. These domain properties may play a role in the distinct trafficking patterns observed during elevated pressures: NHE3 remains in rafts and settles to the base of the microvilli while NaPi2 is freely endocytosed.