Anti-pan ASIC (extracellular) Antibody

Acid-sensing ion channel, Acid-sensing (proton-gated) ion channel, ACCN, ACCN2, ACCN3, ACCN4
    Cat #: ASC-031
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: h, m, r
    Immunogen
    Peptide (C)TRYGKELSMVKIPSK, corresponding to amino acid residues 368 - 382 of rat ASIC1 (Accession P55926); corresponding to amino acid residues 367 – 381 of rat ASIC2 (Accession Q62962); corresponding to amino acid residues 375 – 388 of rat ASIC3 (Accession O35240) and corresponding to amino acid residues 378 – 390 of rat ASIC4 (Accession Q9JHS6). Extracellular loop.
    Accession (Uniprot) Number P55926
    Gene ID 79123
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology ASIC1: Mouse, rat, human – identical.
    ASIC2: Mouse, rat, human – 14/15 amino acid residues identical.
    ASIC3: Mouse, rat, human – 12/15 amino acid residues identical.
    ASIC4: Mouse, rat – 12/15 amino acid residues identical; human – 11/15 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, ih, lci, wb
    May also work in: ifc, ip
    Western blot
    Western blot analysis of rat dorsal root ganglion lysate (lanes 1 and 3) and rat brain membranes (lanes 2 and 4):
    1-2. Anti-pan ASIC (extracellular) Antibody (#ASC-031), (1:500).
    3 -4. Anti-pan ASIC (extracellular) Antibody, preincubated with the control peptide antigen.
    Western blot analysis of mouse brain membranes (lanes 1 and 3) and human SH-SY5Y neuroblastoma cell line lysate (lanes 2 and 4):
    1-2. Anti-pan ASIC (extracellular) Antibody (#ASC-031), (1:500).
    3 -4. Anti-pan ASIC (extracellular) Antibody, preincubated with the control peptide antigen.
    Immunohistochemistry
    Expression of ASIC channels in rat locus coeruleus
    Immunohistochemical staining of perfusion-fixed rat brain frozen sections using Anti-pan ASIC (extracellular) Antibody (#ASC-031), (1:400), followed by goat-anti-rabbit-AlexaFluor-488. ASIC channel staining (green) appears in neuronal soma (arrows). Nuclei are stained with DAPI (blue).
    Live cell imaging / Immunocytochemistry
    Expression of ASIC in rat PC12 cells
    Immunocytochemical staining of live intact rat PC12 pheochromocytoma cells. A. Extracellular staining of cells with Anti-pan ASIC (extracellular) Antibody (#ASC-031), (1:25), followed by goat anti-rabbit-AlexaFluor-488 secondary antibody (green). B. Live view of the cells. C. Merge of A and B.
    References
    1. Poirot, O. et al. (2004) J. Biol. Chem279, 38448.
    2. Krishtal, O. (2003) Trends. Neurosci26, 477.
    3. Brockway, L.M. et al. (2002) Am. J. Physiol. 283, C126.
    4. Ishibashi, K. et al. (1998) Biochem. Biophys. Res. Commun245, 589.
    5. Voilley, N. et al. (2001) J. Neurosci21, 8026.
    6. Wemmie, J.A. et al. (2002) Neuron. 34, 463.
    7. Price, M.P. et al. (2000) Nature. 407, 1007.
    Scientific background

    ASIC channels belong to the ENaC/Degenerin superfamily and are protongated ion channels which are not voltage-dependent. Six different genes encode the different ASIC subunits and splice variants: ASIC1a, ASIC1b, ASIC2a, ASIC2b, ASIC3 and ASIC4.

    ASIC channels sense protons and interact with proteins (namely proteases) and other ions such as Zn2+ and Ca2+ through their extracellular domains. The extracellular loop of ASIC1a undergoes cleavage by extracellular trypsin (a serine protease) which is responsible for shifting the pH dependence of the channel activation and inactivation to more acidic pH1.

    These channels are expressed in the brain and in the nervous system2 but have also been found in the retina, testes, and other tissues3,4 where they are involved in many physiological processes including nociception5, learning and memory6, mechanosensation7 among others.

    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: 29/10/2018

    Alomone Labs is pleased to offer an antibody directed against an epitope of rat ASIC1. The antibody recognizes ASIC1 through ASIC4. Anti-pan ASIC (extracellular) Antibody (#ASC-031) can be used in western blot, immunohistochemistry and immunocytochemistry in living cells. It has been designed to recognize all ASIC channels from rat, human and mouse samples.

    For research purposes only, not for human use
    Citations
    Related Products
      • Products for control experiments
        1. Anti-ASIC1 Antibody (#ASC-014). This antibody recognizes a different epitope and can thus be used as a control for detecting ASIC1.
        2. Anti-ASIC2a Antibody (#ASC-012). This antibody recognizes a different epitope and can thus be used as a control for detecting ASIC2a.
        3. Anti-ASIC3 Antibody (#ASC-018). This antibody recognizes a different epitope and can thus be used as a control for detecting ASIC3.
        4. Anti-ASIC4 Antibody (#ASC-015). This antibody recognizes a different epitope and can thus be used as a control for detecting ASIC4.