Sodium glucose cotransporters (SGLTs) mediate the translocation of carbohydrates across the brush border membrane of different organs such as intestine, kidney and brain. The human SGLT5 (hSGLT5) in particular, is localized in the kidney were it is responsible for mannose and fructose reabsorption from the glomerular filtrate as confirmed by more recent studies on hSGLT5 knockout mice. Here we characterize the functional properties of hSGLT5 expressed in a stable T-Rex- HEK-293 cell line using biochemical and electrophysiological assays. We confirmed that hSGLT5 is a sodium/mannose transporter that is blocked by phlorizin. Li⁺ and H⁺ ions were also able to drive mannose transport. Transport was electrogenic with a 1:1 Na⁺:mannose coupling. Our results moreover indicate that substrates require a pyranose ring with an axial hydroxyl group (-OH) on carbon 2 (C-2). Compared to Na⁺ /glucose cotransport, the level of function of Na⁺/mannose cotransport in rat kidney slices was low.