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Anti-GPR35 (extracellular) Antibody

G-Protein Coupled Receptor 35, Kynurenic Acid Receptor

Cat #: AGR-051
Alternative Name G-Protein Coupled Receptor 35, Kynurenic Acid Receptor
Lyophilized Powder yes
Type: Polyclonal
Host: Rabbit
Reactivity: m, r
Immunogen
  • Peptide (C)EGGF(S)FSSQTRRNFS, corresponding to amino acid residues 156 - 170 of mouse GPR35 (Accession Q9ES90). Extracellular, 2nd loop.
Accession (Uniprot) Number Q9ES90
Gene ID Gpr35
Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
Homology Mouse – 14 out of 15 amino acid residues identical; rat – 12 out of 15 amino acid residues identical. Not recommended for human samples.
Purity Affinity purified on immobilized antigen.
Form Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN3.
Isotype Rabbit IgG
Specificity Not recommended for human samples.
Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
Reconstitution 25 µl, 50 µl or 0.2 ml double distilled water (DDW), depending on the sample size.
Antibody concentration after reconstitution 0.8 mg/ml.
Storage after reconstitution The reconstituted solution can be stored at 4°C for up to 1 week. For longer periods, small aliquots should be stored at -20°C. Avoid multiple freezing and thawing. Centrifuge all antibody preparations before use (10000 x g 5 min).
Preadsorption Control 1 µg peptide per 1 µg antibody.
Standard quality control of each lot Western blot analysis.
Applications: ifc, ih, lci, wb
May also work in: cbe*, ic*, ip*
Western blot
  • Western blot analysis of mouse brain membranes (lanes 1 and 3) and rat brain membranes (lanes 2 and 4):
    Western blot analysis of mouse brain membranes (lanes 1 and 3) and rat brain membranes (lanes 2 and 4):
    1-2. Anti-GPR35 (extracellular) Antibody (#AGR-051), (1:200). 3-4. Anti-GPR35 (extracellular) Antibody, preincubated with GPR35 (extracellular) Blocking Peptide (BLP-GR051).
  • Western blot analysis of rat dorsal root ganglion lysate:
    Western blot analysis of rat dorsal root ganglion lysate:
    1. Anti-GPR35 (extracellular) Antibody (#AGR-051), (1:200). 2. Anti-GPR35 (extracellular) Antibody, preincubated with GPR35 (extracellular) Blocking Peptide (BLP-GR051).
  • Western blot analysis of rat small intestine lysate:
    Western blot analysis of rat small intestine lysate:
    1. Anti-GPR35 (extracellular) Antibody (#AGR-051), (1:200). 2. Anti-GPR35 (extracellular) Antibody, preincubated with GPR35 (extracellular) Blocking Peptide (BLP-GR051).
Immunohistochemistry
  • Expression of GPR35 in rat dorsal root ganglion.
    Expression of GPR35 in rat dorsal root ganglion.
    Immunohistochemical staining of perfusion-fixed frozen rat dorsal root ganglion (DRG) sections with Anti-GPR35 (extracellular) Antibody (#AGR-051), followed by goat anti-rabbit-AlexaFluor-488. A. GPR35 immunoreactivity (green) appeared in neuron profiles (arrows). B. Pre-incubation of the antibody with GPR35 (extracellular) Blocking Peptide (#BLP-GR051), suppressed staining. Cell nuclei are stained with DAPI (blue).
Indirect flow cytometry
  • Cell surface detection of GPR35 by indirect flow cytometry in live intact mouse BV-2 microglia cell line:
    Cell surface detection of GPR35 by indirect flow cytometry in live intact mouse BV-2 microglia cell line:
    ___ Cells.
    ___ Cells + Goat-anti-rabbit-FITC.
    ___ Cells + Anti-GPR35 (extracellular) Antibody (#AGR-051), (5µg) + goat-anti-rabbit-FITC.
References
  1. Yang. et al. (2015), Inflamm. Bowel Dis., 21, 1.
  2. Mackenzie and Milligan. (2017), Neuropharmacology, 113, 661.
  3. Milligan. (2018), Br J Pharmacol., 175, 2543.
  4. Cosi. et al. (2011), Neuropharmacology, 60, 1227.
Scientific background

G- protein coupled receptor 35 (GPR35) is an "orphan" GPCR, whose native ligand is still unknown. It was shown to be involved in the modulation of synaptic transmission, nociception, neuropathic and inflammatory pain, and in various diseases including ulcerative colitis and primary sclerosing cholangitis1 and hence there is a great therapeutic potential in studying its activity.

In vitro, GPR35 is stimulated by high concentrations of a number of small molecules, including kynurenic acid, 2-oleoyllysophosphatidic acid, DHICA, reverse T3, cGMP and it has a wide range of synthetic agonists and antagonists that either activate or inactivate it, respectively (Summarized in2).

 GPR35 is composed of seven trans-membrane domains, and it was shown to share structural homology with various receptors for chemokines; chemotactic cytokines that direct the traffic of leukocytes and other cells in the body. It was shown to be activated by chemokine CXCL17, and hence the role of a chemokine receptor was also assigned to it3.

GPR35 expression has been identified within discrete regions of the central nervous system (CNS) and peripheral nervous system, and lower levels in heart, lung and skeletal muscle, and also in the lower gut and colon and by various white cell groups, including numerous dendritic cell and monocyte populations4.

Application key:

CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, IF- Immunofluorescence, IFC- Indirect flow cytometry, IHC- Immunohistochemistry, IP- Immunoprecipitation, LCI- Live cell imaging, N- Neutralization, WB- Western blot

Species reactivity key:

H- Human, M- Mouse, R- Rat

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Last update: 24/01/2021

Anti-GPR35 (extracellular) Antibody (#AGR-051) is a highly specific antibody directed against an extracellular epitope of the mouse GPR35. The antibody can be used in western blot and live cell flow cytometry applications. It has been designed to recognize GPR35 from mouse and rat samples. The antibody will not recognize the receptor from human samples.

For research purposes only, not for human use
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