pS355/pS356-β2 (phospho-β2-Adrenoceptor Antibody)

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Preview: Agonist-induced Serine355/Serine356 phosphorylation of the β2 Adrenoceptor

Preview: Analysis of time-dependent β2 Adrenoceptor phosphorylation using two phosphosite-specific antibodies

Preview: pS355/pS356-β2 (phospho-β2-Adrenoceptor Antibody)

Agonist-induced Serine355/Serine356 phosphorylation of the β2 Adrenoceptor

Analysis of time-dependent β2 Adrenoceptor phosphorylation using two phosphosite-specific antibodies

pS355/pS356-β2 (phospho-β2-Adrenoceptor Antibody)

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Order number: 7TM0029A
Content: 100 µl
Host: Rabbit

Serine355/Serine356 (S355/S356) is a major phosphorylation site of the β2 adrenoceptor.... more

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Serine355/Serine356 (S355/S356) is a major phosphorylation site of the β2 adrenoceptor. The pS355/pS356-β2 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation. S355/S356 phosphorylation is primarily mediated by GRK6 and is a key regulator of β2 desensitization, β-arrestin recruitment and internalization.

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Alternative Names β2, B2AR, ADRB2R, β2 Adrenergic... more

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Alternative Names	β2, B2AR, ADRB2R, β2 Adrenergic Receptor, β2-Adrenoceptor
IUPHAR Target ID	29
UniProt ID	P07550
Western Blot (WB)	1:1000
Immunocytochemistry (ICC)	1:100
Species Reactivity	Human
Host / Isotype	Rabbit / IgG
Class	Polyclonal
Immunogen	A synthetic phosphopeptide derived from human β2 around the phosphorylation site of Ser355/Ser356.
Form	Liquid
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 Serine355/Serine356 phosphorylation of the β2-Adrenoceptor.... more

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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. Analysis of time-dependent β2-Adrenoceptor phosphorylation using two phosphosite-specific antibodies. Upper two panels, HEK293 cells stably expressing the β2-Adrenoceptor (β2) were exposed to 1µM Isoprenaline for increasing periods of time ranging from 5 to 120 s. Cells were lysed and immunoblotted with the anti-pS355/pS356-β2 antibody (7TM0029A) or 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. Immunocytochemical identification of Serine355/Serine356 phosphorylation of β2-Adrenoceptor. Hela cells transiently expressing the FLAG-tagged β2-Adrenoceptor (β2) were exposed to 10 μM of specific β2-adrenoceptor agonist Isoprenaline for 5 min and immunocytochemically stained with an anti-FLAG antibody or the anti-pS355/pS356-β2 antibody (7TM0029A) at a dilution of 1:100.

Benkel T, Zimmermann M, Zeiner J, Bravo S, Merten N, Lim VJY, Matthees ESF, Drube J,... more

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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.

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