Figure 2

Functional characterization of β2AR-T4L. (A) The protein was expressed in vitro and ligand binding was assessed on crude material as described in the text. Protein yield was estimated by trace-labeling with [³⁵S]Met. As a control, purified receptor expressed in insect cells [5] was used. (B). Binding assays for (A) and (B) were performed as described in methods [5]. Propranolol was used as the unlabeled competitor. Experiments in (A) were performed using The MembraneMax Protein Expression Kit (Life Technologies, Carlsbad, CA) using DTT-free buffers with the indicated modifications (see methods). Kd and Bmax values were calculated using GraphPad PRISM software (GraphPad Software, San Diego, CA). Abbreviations: NSB, non-specific binding;