Western Blot Analysis of Insect Odorant Receptors

Expression of ORs by western blot analysis was essentially as previously described (Tsitoura et al., 2010 (link)). In particular, Myc-tagged ORs 1, 2, 9, and 53 were detected in total lysates of transiently transfected Hi5 cells, by the use of mouse anti-Myc antibody (Cell Signaling 9B11, 1:1,000 dilution). For Flag-tagged ORco protein detection in cells expressing either ORco (fORco) alone or its combinations with various ORx subunits, the anti-Flag antibody (Sigma F1804, 1:800 dilution) was used. In the latter case, Hi5 cells were transfected with constructs directing expression of fORco, OR1/fORco, OR2/fORco, OR9/fORco or OR53/fORco and Photina, and cells were used for western blot analysis and, in the presence of co-expressed Photina, functional assays.