Anti-VPAC1 (extracellular) Antibody

VIP and PACAP receptor 1, Vasoactive intestinal polypeptide receptor 1, VIPR1, Pituitary adenylate cyclase activating polypeptide receptor type II, PACAP type II receptor
    Cat #: AVR-001
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: h, m, r
    Immunogen
    Peptide (C)EEAQLENETIG(S)SK, corresponding to amino acid residues 52-65 of human VPAC1 (Accession P32241). Extracellular, N-terminus.
    Accession (Uniprot) Number P32241
    Gene ID 7433
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Human - 13/14 amino acid residues identical; rat, mouse - 12/14 amino acid residues identical.
    Purity Affinity purified on immobilized antigen.
    Formulation Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN3.
    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).
    Control antigen storage before reconstitution Lyophilized powder can be stored intact at room temperature for 2 weeks. For longer periods, it should be stored at -20°C.
    Control antigen reconstitution 100 µl double distilled water (DDW).
    Control antigen storage after reconstitution -20ºC.
    Preadsorption Control 1 µg peptide per 1 µg antibody.
    Standard quality control of each lot Western blot analysis.
    Applications: ic, ifc, ih, lci, wb
    May also work in: ip
    Western blot
    Western blot analysis of rat brain lysate (lanes 1 and 4), mouse brain membranes (lanes 2 and 5) and human Jurkat T cell leukemia cell lysate (lanes 3 and 6):
    1-3. Anti-VPAC1 (extracellular) Antibody (#AVR-001), (1:400).
    4-6. Anti-VPAC1 (extracellular) Antibody, preincubated with the control peptide antigen.
    Immunohistochemistry
    Expression of VPAC1 in rat amygdala
    Immunohistochemical staining of immersion-fixed, free floating rat brain frozen sections using Anti-VPAC1 (extracellular) Antibody (#AVR-001), (1:100). A. VPAC1 staining (red) is apparent in basolateral amygdala neurons (horizontal arrow). B. Cell nuclei in the same section are stained with DAPI (Blue). C. Merge of the two images.
    Indirect flow cytometry
    Indirect flow cytometry analysis of live intact Jurkat (human T cell leukemia cells) cell line:
    ___ Cells + goat-anti-rabbit-DyLight-488.
    ___ Cells + Anti-VPAC1 (extracellular) Antibody (#AVR-001), (1:20) + goat-anti-rabbit-DyLight-488.
    The control antigen is not suitable for this application.
    Live cell imaging / Immunocytochemistry
    Expression of VPAC1 in human HT-29 cells
    Immunocytochemical staining of live intact human HT-29 colorectal adenocarcinoma cells. A. Extracellular staining of live cells with Anti-VPAC1 (extracellular) Antibody (#AVR-001), (1:25), followed by goat anti-rabbit-AlexaFluor-594 secondary antibody (red). B. Cell nuclei were visualized using Hoechst 33342 (blue). C. Live view of the cells.
    References
    1. Vaudry, D. et al. (2000) Pharm. Rev. 52, 269.
    2. Gozes, I. et al. (2003) Curr Pharm Des. 9, 483.
    3. Laburthe, M. et al. (2002) Receptors Channels 8, 137.
    4. Laburthe, M. et al. (1996) Ann. N. Y. Acad. Sci. 805, 94.
    5. Usdin, T. et al. (1994) Endocrinology 135, 2662.
    6. Reubi, J.C. et al. (2000). Ann. N. Y. Acad. Sci. 921, 1.
    7. Said, S.I. (1991) Trends Endocrinol. Metab. 2, 107.
    Scientific background

    Vasointestinal peptide (VIP) and pituitary adenylate cyclase–activating peptide (PACAP) belong to the glucagon hormone superfamily, which includes secretin, growth hormone–releasing hormone (GHRH), glucagon, glucagon-like peptides 1 and 2 (GLP-1 and GLP-2), peptide histidine methionine (PHM), and glucose-dependent insulinotropic polypeptide (GIP)1. PACAP and VIP effects have been described in the digestive tract, cardiovascular system, airways, reproductive system, immune system, endocrine glands, and brain2. VIP and PACAP share a common G-protein coupled receptor, VPAC13.

    VPAC1 is a membrane-associated protein and shares significant homology with members of the G-protein coupled class B receptor family, the most important of which is the presence of large N-terminal extracellular domains which contain 10 highly conserved amino acids including six cysteines, putative N-terminal leader sequences and several potential N-glycosylation sites4. In the CNS, VPAC1 receptors are abundantly localized in piriform cortex, cerebral cortex, suprachiasmatic nucleus, hippocampus, and pineal gland5. In peripheral tissues, VPAC1 receptors have been found in breast, kidney, liver, lung, prostate, spleen, and mucosa of the gastrointestinal tract6. VPAC1 mediates a large array of VIP and PACAP actions on exocrine secretion, hormones release, muscle relaxation, metabolism, fetus growth, tumor cells and embryonic brain development7.

    Application key:

    CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, 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
    Last update: 30/10/2018

    Anti-VPAC1 (extracellular) Antibody (#AVR-001) is a highly specific antibody directed against an epitope of the human protein. The antibody can be used in western blot, immunohistochemistry, immunocytochemistry, live cell imaging, and indirect flow cytometry applications. It has been designed to recognize VPAC1 from mouse, rat, and human samples.

    For research purposes only, not for human use
    Citations
    Western blot citations
    1. Mouse carotid lysate (1:800).
      Ivic, I. et al. (2017) J. Vasc. Res. 54, 180.
    Related Products
      • Antibodies for live cell flow cytometry experiments
        1. Anti-VPAC1 (extracellular)-FITC Antibody (#AVR-001-F). This FITC-conjugated antibody can be used to detect Ghrelin receptor in live cell flow cytometry.