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- Order number: 7TM0239A-IC
- Content: 300 µl
- Host: Rabbit
Serine594/Threonine597/Serine599 (S362/S363/S364) is major GRK2 phosphorylation site of the SMO receptor. The pS594/pT597/pS599-SMO antibody detects phosphorylation in response to agonists. The pS594/pT597/pS599-SMO antibody can be used for detection of the subcellular location of phosphorylated SMO by immunocytochemistry or by immunohistochemistry in mouse brain.
| Alternative Names | SMOH | FZD11 | frizzled family member 11 | bnb | smoothened, frizzled class receptor | smoothened |
| IUPHAR Target ID | 293 |
| UniProt ID | Q99835 (human), P56726 (mouse), P97698 (rat) |
| Western Blot (WB) | 1:1000 |
| Immunocytochemistry (ICC) | 1:1000 |
| Immunohistochemistry (IHC) | 1:1000 |
| Species Reactivity | Human, Mouse, Rat |
| Host / Isotype | Rabbit / IgG |
| Class | Polyclonal |
| Immunogen | A synthetic phosphopeptide derived from mouse SMO around the phosphorylation site of Ser594/Thr597/Ser599. |
| 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. Immunohistochemical Identification of Serine594/Threonine597/Serine599 phosphorylation of the SMO Receptor in primary cilia in the mouse neural tube. Neural tubes from wild-type E9.5 mice were stained for pS594/pT597/pS599-SMO antibody and Arl13b. Higher magnification images were acquired in the ventral neural tube region, the area with the highest activation of the Hh pathway.
Figure 2. Immunocytochemical identification of phospho-SMO Receptor in NIH3T3 cells. NIH3T3 cells, stably expressing the Flag-tagged SMO receptor, were either not exposed or exposed to 1 µM SAG21K overnight and immunocytochemically stained with anti-pS594/pT597/pS599-SMO antibody (7TM0239A-IC) at a dilution of 1:1000. Note, phospho-SMO receptors were detected primary clilia only in agonist-treated cells.
Figure 3. Agonist-induced Serine594/Threonine597/Serine599 phosphorylation of the SMO Receptor. Upper panel, NIH3T3 cells stably expressing the Flag-tagged SMO receptor were either not exposed or exposed to 1 µM SAG21K for 4 hours in the absence or presence of 30 µM of the the GRK2/3 inhbibitor Compound 101. Cells were lysed and immunoblotted with anti-pS594/pT597/pS599-SMO antibody (7TM0239A-IC) at a dilution of 1:1000. Lower panel, blot was probed with the phosphorylation-independent anti-FLAG antibody (7TM00FLAG) at a dilution of 1:1000 to confirm equal amounts of total SMO in each sample.
Walker MF, Zhang J, Steiner W, Ku PI, Zhu JF, Michaelson Z, Yen YC, Long AB, Casey MJ, Poddar A, Nelson IB, Arveseth CD, Nagel F, Clough R, LaPotin S, Kwan KM, Schulz S, Stewart RA, Tesmer JJG, Caspary T, Subramanian R, Ge X, Myers BR. A Ciliary SMOOTHENED-GRK2-PKA Signaling Pathway Initiates Hedgehog Signal Transduction. bioRxiv. 2023 May 11:2023.05.10.540226. doi: 10.1101/2023.05.10.540226. PMID: 37214942; PMCID: PMC10197709.
