NOP (non-phospho), Nociceptin/Orphanin FQ Receptor Antibody

Figure 1. Validation of the Nociceptin/Orphanin FQ Receptor in transfected HEK293 cells. Native HEK293 cells (MOCK) or HEK293 cells stably expressing the Nociceptin/Orphanin FQ Receptor (NOP) were lysed and immunoblotted with the phosphorylation-independent anti-NOP antibody (7TM0320N-WB) at a dilution of 1:1000.

Figure 2. Analysis of dose-dependent Nociceptin/Orphanin FQ Receptor phosphorylation in vivo in NOP-eGFP knock-in mice using two phosphosite-specific antibodies. Upper two panels, NOP-eGFP knock-in mice stably expressing the Nociceptin/Orphanin FQ Receptor tagged with enhanced green fluorescent protein (NOP-eGFP) were intraperitoneal injected with 0.9% NaCl or increasing concentrations of specific Nociceptin/Orphanin FQ receptor agonist AT-202 ranging from 0.3 to 30 mg/kg for 30 minutes. Mice were euthanized and brains were removed. The NOP-eGFP receptor was immunoprecipitated with anti-GFP protein agarose beads and immunoblotted with the anti-pS351-NOP antibody (7TM0320B) or anti-pT362/pS363-NOP antibody (7TM0320C) at a dilution of 1:1000. Lower panel, blot was stripped and reprobed with the phosphorylation-independent anti-NOP antibody (7TM0320N-WB) at a dilution of 1:1000 to confirm equal loading of the gel.

Mann A, Moulédous L, Froment C, O'Neill PR, Dasgupta P, Günther T, Brunori G, Kieffer BL, Toll L, Bruchas MR, Zaveri NT, Schulz S. Agonist-selective NOP receptor phosphorylation correlates in vitro and in vivo and reveals differential post-activation signaling by chemically diverse agonists. Sci Signal. 2019 Mar 26;12(574). pii: eaau8072. doi: 10.1126/scisignal.aau8072. PubMed PMID: 30914485; PubMed Central PMCID: PMC6934085.