Immobilized protein has advanced in many areas like drug discovery. While this field evolved rapidly over the last three decades, the immobilization platform for the G-protein-coupled receptor (GPCR) remains unpromising due to its instability under the relatively harsh conditions of current methodologies. Taking beta2-adrenoceptor (β2-AR) as an example, we presented here a general strategy for immobilization of GPCRs by combining the His6-tag trap system, conformation-specific aptamer, and target binding induced DNA hybridization. Morphology characterization by diverse assays confirmed a monolayer of β2-AR on the microsphere surface. The radio-ligand binding assay and immuno-transmission electron microscopy showed desirable ligand- and antibody-binding activities. A case study of chromatography using the immobilized receptor as a stationary phase exhibited a demonstrable conformation specificity that enables the selective recognition of the receptor agonists or antagonists. Owing to the competitive strand displacement during the immobilization, the method proved to be capable of sensitively and directly determining the receptor density on the surface which enormously challenges most of the reported assays. This method is possible to turn into a general strategy for the immobilization of GPCRs with a defined orientation, conformation, function, and density, thus paving the way for precisely realizing the receptor-ligand binding interaction and screening the receptor agonist or antagonist with high efficiency.