β2-Adrenoceptor Sample Pack (phospho- and non-phospho-β2-Adrenoceptor Antibodies)

Figure 1. Agonist-induced Serine355/Serine356 phosphorylation of the β2-Adrenoceptor. Upper panel, HEK293 cells stably expressing the β2-Adrenoceptor (β2) were either not exposed or exposed to 1 µM of specific β2-adrenoceptor agonist Isoprenaline or 0.1 μM PMA (Phorbol 12-Myristate 13-Acetate) for 30 minutes. Cells were lysed and immunoblotted with the anti-pS355/pS356-β2 antibody (7TM0029A) at a dilution of 1:1000. Lower panel, blot was stripped and reprobed with the phosphorylation-independent anti-β2 antibody (7TM0029N) at a dilution of 1:1000 to confirm equal loading of the gel.

Figure 2. Agonist-induced Threonine360/Serine364 phosphorylation of the β2-Adrenoceptor. Upper panel, HEK293 cells stably expressing the β2-Adrenoceptor (β2) were either not exposed or exposed to 1 µM of specific β2-adrenoceptor agonist Isoprenaline or 0.1 μM PMA (Phorbol 12-Myristate 13-Acetate) for 30 minutes. Cells were lysed and immunoblotted with the anti-pT360/pS364-β2 antibody (7TM0029B) at a dilution of 1:1000. Lower panel, blot was stripped and reprobed with the phosphorylation-independent anti-β2 antibody (7TM0029N) at a dilution of 1:1000 to confirm equal loading of the gel.

Figure 3. Agonist-induced Serine261/Serine262 phosphorylation of the β2-Adrenoceptor. Upper panel, HEK293 cells stably expressing the β2-Adrenoceptor (β2) were either not exposed or exposed to 1 µM of specific β2-adrenoceptor agonist Isoprenaline for 30 minutes. Cells were lysed and immunoblotted with the anti-pS261/pS262-β2 antibody (7TM0029G) at a dilution of 1:1000. Lower panel, blot was stripped and reprobed with the phosphorylation-independent anti-β2 antibody (7TM0029N) at a dilution of 1:1000 to confirm equal loading of the gel.

Figure 4. Validation of the β2-Adrenoceptor in transfected HEK293 cells. Native HEK293 cells (MOCK) or HEK293 cells stably expressing the β2-Adrenoceptor (β2) were lysed and immunoblotted with the phosphorylation-independent anti-β2 antibody (7TM0029N-WB) at a dilution of 1:1000.

Benkel T, Zimmermann M, Zeiner J, Bravo S, Merten N, Lim VJY, Matthees ESF, Drube J, Miess-Tanneberg E, Malan D, Szpakowska M, Monteleone S, Grimes J, Koszegi Z, Lanoiselée Y, O'Brien S, Pavlaki N, Dobberstein N, Inoue A, Nikolaev V, Calebiro D, Chevigné A, Sasse P, Schulz S, Hoffmann C, Kolb P, Waldhoer M, Simon K, Gomeza J, Kostenis E. How Carvedilol activates β2-adrenoceptors. Nat Commun. 2022 Nov 19;13(1):7109. doi: 10.1038/s41467-022-34765-w. PMID: 36402762; PMCID: PMC9675828.

Burghi V, Paradis JS, Officer A, Adame-Garcia SR, Wu X, Matthees ESF, Barsi-Rhyne B, Ramms DJ, Clubb L, Acosta M, Tamayo P, Bouvier M, Inoue A, von Zastrow
M, Hoffmann C, Gutkind JS. Gαs is dispensable for β-arrestin coupling but dictates GRK selectivity and is predominant for gene expression regulation by
β2-adrenergic receptor. J Biol Chem. 2023 Nov;299(11):105293. doi:10.1016/j.jbc.2023.105293. Epub 2023 Sep 27. PMID: 37774973; PMCID: PMC10641165.

Radoux-Mergault A, Oberhauser L, Aureli S, Gervasio FL, Stoeber M.Subcellular location defines GPCR signal transduction. Sci Adv. 2023 Apr 21;9(16):eadf6059. doi: 10.1126/sciadv.adf6059. Epub 2023 Apr 19. PMID: 37075112; PMCID: PMC10115417.