Regulation of transport processes in renal epithelial cells in response to alpha 2-adrenergic receptor (alpha 2-AR) stimulation is likely due to specific effects of the A-, B-, or C-subtype alpha 2-AR. The purpose of the present study was 1) to examine the precise localization, using subtype-specific antibodies, the B-subtype alpha 2-AR protein, the most abundant alpha 2-AR in rat kidney; and 2) to determine the effect of dietary potassium restriction, a factor known to increase alpha 2B-ARs, on the distribution of receptors. We employed a method to generate a specific antibody to the third intracellular loop of the alpha 2B-AR and demonstrated the specificity of this antibody. Our results demonstrated, for the first time, evidence for the alpha 2B-receptor protein in the basolateral membrane of proximal convoluted and straight tubules. No specific immunoreactivity was detected in other nephron segments. Finally, the increase in alpha 2B-AR expression observed previously was due to the recruitment of alpha 2B-ARs to the basolateral membrane of proximal tubules.