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- Order number: 7TM0016D
- Content: 100 µl
- Host: Rabbit
Serine298/Serine300 is a major phosphorylation site of the M4 Muscarinic Acetylcholine Receptor. The pS298/pS300-M4 antibody detects phosphorylation in response to high-efficacy agonists but not after PKC activation. S298/S300 phosphorylation is a key regulator of M4 desensitization, β-arrestin recruitment and internalization.
| Alternative Names | HM3, Chrm 4, Cholinergic Receptor muscarinic 4 |
| IUPHAR Target ID | 16 |
| UniProt ID | P08173 |
| Western Blot (WB) | 1:1000 |
| Immunocytochemistry (ICC) | - |
| Species Reactivity | Human |
| Host / Isotype | Rabbit / IgG |
| Class | Polyclonal |
| Immunogen | A synthetic phosphopeptide derived from human M4 around the phosphorylation site of Thr315/Thr316. |
| 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 Serine298/Serine300 phosphorylation of the M4 Muscarinic Acetylcholine Receptor. Upper panel, HEK293 cells stably expressing the M4 Muscarinic Acetylcholine Receptor (M4) and high level of GRK2 were either not exposed or exposed to 100 µM Carbachol for 30 minutes. Cells were lysed and immunoblotted according to 7TM Western Blot Protocol with the anti-pS298/pS300-M4 antibody (7TM0016D) at a dilution of 1:1000. Lower panel, blot was stripped and reprobed with the phosphorylation-independent anti-M4 antibody (7TM0016N) to confirm equal loading of the gel.
Figure 2. Immunohistochemical identification of Serine298/Serine300 phosphorylation of M4 Receptor in striatum. Sections were dewaxed, microwaved in citric acid, and incubated with anti-pS298/pS300-M4 (M4 Receptor) antibody (7TM0016D) at a dilution of 1:100. Sections were then sequentially treated with biotinylated anti-rabbit IgG and avidin-biotin solution. Sections were then developed in 3,3-diaminobenzidine (DAB)-glucose oxidase and lightly counterstained with hematoxylin.
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