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- Order number: 7TM0226A-IC
- Content: 300 µl
- Host: Rabbit
Serine296/Serine297 (S296/S297) is major phosphorylation site of the FFA Receptor 2 (FFA2). The pS296/pS297-FFA2 antibody detects constitutive phosphorylation of FFA2.
Alternative Names | FFAR2, Free Fatty Acid Receptor 2, GPCR3 |
IUPHAR Target ID | 226 |
UniProt ID | O15552 |
Western Blot (WB) | 1:1000 |
Immunocytochemisty (ICC) | 1:200 |
Immunohistochemistry (IHC) | 1:100 |
Species Reactivity | Human |
Host / Isotype | Rabbit / IgG |
Class | Polyclonal |
Immunogen | A synthetic phosphopeptide derived from human FFA2 around the phosphorylation site of Ser296/Ser297. |
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 Serine296/Serine297 phosphorylation of the FFA Receptor 2 in mouse white adipose tissue and peyer's patches. Sections from transgenic hFFAR2- DREADD- HA- expressing mouse line were treated with 100 µM 4- methoxy- 3- methyl- benzoic acid (MOMBA), dewaxed, microwaved in citric acid, and incubated with anti-pS425/pS429-VPAC2 antibody (7TM0372C-IC) or non-phospho anti-VPAC2 antibody (7TM0372N-IC) at a dilution of 1:100. Sections were then sequentially treated with biotinylated anti-rabbit IgG and avidin-biotin solution.Color was developed by incubation in 3-amino-9-ethylcarbazole (AEC), and sections were counterstained with hematoxylin.
Figure 2. Immunocytochemical identifiction of Serine296/Serine297 phosphorylation of the FFA Receptor 2. Flp-In TREx 293 cells stably expressing human FFA2-HA were either treated with vehicle or 2mM C3 for 2 min and immunocytochemically stained with the anti-pS296/pS297-FFA2 antibody (7TM0226A-IC) at a dilution of 1:200.
Figure 3. Constitutive Serine296/Serine297 phosphorylation of the FFA Receptor 2. Upper panel, parental or Flp-In TREx 293 harbouring human FFA2-eYFP induced to express the receptor construct (+ dox) were treated with vehicle or 2mM propanoic acid (C3) for 5 min. Cells were lysed and receptor construct enriched with a GFP-trap, before immunoblotting with the anti-pS296/pS297-FFA2 antibody (7TM0226A-IC) at a dilution of 1:1000. Lower panel, blot was stripped and reprobed with a phosphorylation-independent anti-GFP antibody to confirm equal loading of the gel.
Barki N, Jenkins L, Marsango S, Dedeo D, Bolognini D, Dwomoh L, Abdelmalik AM, Nilsen M, Stoffels M, Nagel F, Schulz S, Tobin AB, Milligan G. Phosphorylation bar-coding of free fatty acid receptor 2 is generated in a tissue-specific manner. Elife. 2023 Dec 12;12:RP91861. doi: 10.7554/eLife.91861. PMID: 38085667; PMCID: PMC10715726.
Ruhnke N, Beyer AL, Kaemmerer D, Sänger J, Schulz S, Lupp A. Expression of free fatty acid receptor 2 in normal and neoplastic tissues. Exp Mol Pathol. 2024 Jun;137:104902. doi: 10.1016/j.yexmp.2024.104902. Epub 2024 May 23. PMID: 38788249.