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pT306/pT310-FFA2 (IHC grade phospho-FFA2 Antibody)

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  • 7TM0226B-IC
  • 300 µl
  • Rabbit
Threonine306/Threonine310 (T306/T310) is major phosphorylation site of the FFA 2 Receptor... more

Threonine306/Threonine310 (T306/T310) is major phosphorylation site of the FFA 2 Receptor (FFA2). The pT306/pT310-FFA2 antibody detects phosphorylation in response to agonists. T306/T310 phosphorylation is likely to be involved in efficient ligand sequestration by FFA2.

  Alternative Names FFAR2, Free Fatty Acid Receptor 2, GPCR3 IUPHAR... more

 

Alternative Names FFAR2, Free Fatty Acid Receptor 2, GPCR3
IUPHAR Target ID  226
UniProt ID  O15552
Western Blot (WB)  1:1000
Immunocytochemistry (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 Thr306/Thr310.
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 Threonine306/Threonine310 phosphorylation of... more

Figure 1. Immunohistochemical Identification of Threonine306/Threonine310 phosphorylation of the FFA Receptor 2 in mouse neuroendocrine cells 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), fixed, dewaxed, microwaved in citric acid, and incubated with anti-pT306pT310-FFA2 antibody (7TM0226B-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 Threonine306/Threonine310 phosphorylation of the FFA Receptor 2. Flp-In TREx 293 cells stably expressing hFFA2-HA were either treated with vehicle or 2 mM C3 for 2 min and immunocytochemically stained with the anti-pT306pT310-FFA2 antibody (7TM0226B-IC) at a dilution of 1:200. Note, Threonine306/Threonine310-phosphorylated FFA2 receptors were not detectable in untreated cells (vehicle). Threonine306/Threonine310-phosphorylated FFA2 receptors were seen at the plasma membrane after 2 min C3-treatment.

Figure 3. Agonist-induced Threonine306/Threonine310 phosphorylation of the FFA2 Receptor. 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-pT306/pT310-FFA2 antibody (7TM0226B-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,... more

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.

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