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- Order number: 7TM0029B
- Content: 100 µl
- Host: Rabbit
Threonine360/Serine364 (T360/S364) is a major phosphorylation site of the β2 adrenoceptor. The pT360/pS364-β2 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation. T360/S364 phosphorylation is primarily mediated by GRK2 and is a key regulator of β2 desensitization, β-arrestin recruitment and internalization.
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 Thr360/Ser364. |
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 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 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 Threonine360/Serine364 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-pT360/pS364-β2 antibody (7TM0029B) at a dilution of 1:100.
Figure 4. Immunocytochemical identification of Threonine360/Serine364 phosphorylation of β2-Adrenoceptor. Hela cells transiently expressing the β2-Adrenoceptor (β2) were either not exposed or exposed to 10 μM of specific β2-adrenoceptor agonist Isoprenaline and immunocytochemically stained with the anti-pT360/pS364-β2 antibody (7TM0029B) at a dilution of 1:100. Note, Threonine360/Serine364-phosphorylated β2 receptors were not detectable in untreated cells (0 min). Threonine360-phosphorylated β2 receptors were seen at the plasma membrane after 5 min Isoprenaline stimulation.
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