The cDNAs encoding for three subtypes of adrenergic receptors, alpha(1A)-, alpha(1B)- and alpha(1D)-ARs, were cloned and expressed in HEK 293 cells. Expression of alpha(1A)- and alpha(1B)-AR subtypes in HEK 293 cells was stable even with increased passages but that of alpha(1D)-AR was not. Cellular localization studies using immunofluorescence and flow cytometry revealed that expression of alpha(1A)- and alpha(1B)-ARs was primarily localized on the cell membrane whereas expression of alpha(1D)-AR was predominantly intracellular. Our studies clearly demonstrated that the culturing of the recombinant cell lines expressing alpha(1D)-AR in charcoal/dextran treated fetal bovine serum (FBS) resulted in targeting of alpha(1D)-AR to the cell membrane and thus, significantly improving its stability and availability for ligand binding studies.