pT427/pS429-GPR75 (IHC grade phospho-GPR75 Antibody)

Figure 1. Immunohistochemical identification of Threonine 427/Serine429 phosphorylation of G-protein coupled Receptor 75 in mouse brain. Sections were dewaxed, microwaved in citric acid, and incubated with anti-pT427/pS429-GPR75) antibody (7TM0115B-IC) at a dilution of 1:100. Sections were then sequentially treated with biotinylated anti-rabbit IgG and avidin-biotin solution.Color was developed by incubation in 3-amino-9-ethylcarbazole (AEC), and sections were counterstained with hematoxylin.

Figure 2. Agonist-induced Threonine427/Serine429 phosphorylation of the G-protein coupled Receptor 75. Upper panel, HEK293 cells stably expressing the G-protein coupled receptor 75 (GPR75) are marked as wildtype (WT). HEK293 cells stably expressing the G-protein coupled receptor 75 (GPR75) and have full knock-out of G protein-coupled receptor kinases (GRK) 2,3,4 and 5 are marked as ΔGRK2/3/4/5. Knock-out cells were treated or not treated with 0.1 μM PMA (Phorbol 12-Myristate 13-Acetate) or 1 µM Forskolin. Cells were lysed and immunoblotted with the anti-pT427/pS429-GPR75 antibody (7TM0115B-IC) at a dilution of 1:1000. Lower panel, blot was stripped and reprobed with an phosphorylation-independent anti-GPR75 antibody to confirm equal loading of the gel.

Divorty N, Jenkins L, Ganguly A, Butcher AJ, Hudson BD, Schulz S, Tobin AB, Nicklin SA, Milligan G. Agonist-induced phosphorylation of orthologues of the orphan receptor GPR35 functions as an activation sensor. J Biol Chem. 2022 Mar;298(3):101655. doi: 10.1016/j.jbc.2022.101655. Epub 2022 Jan 29. PMID: 35101446; PMCID: PMC8892012.