Threonine361 (T361) is a major phosphorylation site of the delta-opioid receptor (DOP). The pT361-DOP antibody detects phosphorylation in response to agonists but not after PKC activation. T361 phosphorylation is a key regulator of DOP desensitization, β-arrestin recruitment and internalization. The pT361-DOP antibody can detect phosphorylated DOP in mouse brain in vivo.
|Alternative Names||DOP, OPRD1, δ-Opioid Receptor|
|IUPHAR Target ID||317|
|UniProt ID||P41143 (human) P32300 (mouse) P33533 (rat)|
|Western Blot (WB)||1:1000|
|Species Reactivity||Human, Mouse, Rat|
|Host / Isotype||Rabbit / IgG|
|Immunogen||A synthetic phosphopeptide derived from human DOP around the phosphorylation site of Thr361|
|Purification||Antigen affinity chromatography|
|Storage buffer||Dulbecco's PBS, pH 7.4, with 150 mM NaCl, 0.02% sodium azide|
|Storage conditions||short-term 4°C, long-term -20°C|
Figure 1. Agonist-induced Threonine361 phosphorylation of the δ-Opioid Receptor. Upper panel, HEK293 cells stably expressing the δ-Opioid Receptor (DOP) were either not exposed or exposed to 10 μM DPDPE ([D-Pen2,D-Pen5]-enkephalin) or 0.1 μM PMA (Phorbol 12-Myristate 13-Acetate) for 30 minutes. Cells were lysed and immunoblotted with the anti-pT361-DOP antibody (7TM0317A) at a dilution of 1:1000. Lower panel, blot was stripped and reprobed with a phosphorylation-independent anti-DOP antibody to confirm equal loading of the gel.
Figure 2. Analysis of dose-dependent δ-Opioid Receptor phosphorylation using two phosphosite-specific antibodies. Upper two panels, HEK293 cells stably expressing the δ-Opioid Receptor (DOP) were either not exposed or exposed to increasing concentrations of selective δ-opioid receptor agonist SNC80 ranging from 1 nM to 10 μM for 30 minutes. Cells were lysed and immunoblotted with the anti-pT361-DOP antibody (7TM0317A) or anti-pS363-DOP antibody (7TM0317B) at a dilution of 1:1000. Lower panel, blot was stripped and reprobed with a phosphorylation-independent anti-DOP antibody to confirm equal loading of the gel.
Mann A, Liebetrau S, Klima M, Dasgupta P, Massotte D, Schulz S. Agonist-induced phosphorylation bar code and differential post-activation signaling of the delta opioid receptor revealed by phosphosite-specific antibodies. Sci Rep. 2020 May 22;10(1):8585. doi: 10.1038/s41598-020-65589-7. PMID: 32444688