Anti-ASIC3 Antibody

Acid-sensing ion channel 3, Amiloride-sensitive cation channel 3, ACCN3, DRASIC
    Cat #: ASC-018
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: h, m, r
    Immunogen
    Peptide KPRSGLEEAQRRQAS(C), corresponding to amino acid residues 2-16 of rat ASIC3 (Accession O35240). Intracellular, N-terminus.
    Accession (Uniprot) Number O35240
    Gene ID 286920
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Mouse - 14/15 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 -20°C.
    Preadsorption Control 1 µg peptide per 1 µg antibody.
    Standard quality control of each lot Western blot analysis.
    Applications: ic, ih, wb
    May also work in: ifc, ip
    Western blot
    Western blot analysis of rat DRG lysate:
    1. Anti-ASIC3 Antibody (#ASC-018), (1:200).
    2. Anti-ASIC3 Antibody, preincubated with the control peptide antigen.
    Immunohistochemistry
    Immuno-colocalization of ASIC1 and ASIC3 in rat DRG
    Immunohistochemical staining of perfusion-fixed frozen rat dorsal root ganglia (DRG) sections using Guinea pig Anti-ASIC1 Antibody (#AGP-053), (1:400) and rabbit Anti-ASIC3 Antibody (#ASC-018), (1:400). A. ASIC3 staining. B. ASIC1 staining in same section (green). C. Merge of the two images shows some co-localization (arrow). Cell nuclei were stained with DAPI (blue).
    Rat trigeminal ganglia (1:400) (Yan, J. et al. (2013) Headache 53, 1250.).
    Immunocytochemistry
    Human SH-SY5Y cells (Xiong, Q.J. et al. (2012) Am. J. Physiol. 303, C376.)
    References
    1. Kellenberger, S. and Schild, L. (2002) Physiol. Rev. 82, 735.
    2. Chen, C.C. et al. (2002) Proc. Natl. Acad. Sci. U.S.A99, 8992.
    3. Price, M.P. et al. (2001) Neuron 32, 1071.
    Scientific background

    ASIC3 is a member of a family of Na+ channels that are activated by external protons. The family includes another three members ASIC1ASIC2 and ASIC4. The ASICs are in fact part of a larger superfamily termed degenerin/epithelial Na+ channels (DEG/ENaC) and share with it the same basic characteristics: two transmembrane spanning domains, a large extracellular domain and short intracellular N and C termini.

    The functional channel is composed of 4 subunits that can be assembled as homo or heterotetramers with the other ASIC subunits.

    A drop in external pH opens the channel resulting in an inward cation current that is quickly inactivated even in the continuous presence of protons in the medium, although a small residual current may persist.

    Several lines of evidence indicate that ASIC3 may function as a pain sensor. First, it is specifically located in dorsal root ganglia (DRG) neurons and in nociceptors that are involved in pain detection and transmission. In addition, in conditions such as inflammation and cardiac ischemia that include tissue acidosis and thus pain, ASIC3 currents have been detected. Finally, ASIC3 deficient mice display altered sensitivity to high intensity pain produced by heat or acid.

    Interestingly, while in rats ASIC3 is expressed almost exclusively in DRG neurons, in humans its expression is more widespread suggesting a more extensive role in human nociception.

    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
    Image & Title:

    Anti-ASIC3 Antibody
    Expression of ASIC3 in rat trigeminal ganglia.Immunohistochemical staining of rat trigeminal ganglia (TG) sections using Anti-ASIC3 Antibody (#ASC-018). ASIC3 staining (green) appears in TG neurons (left panel) and coincides with FAST DiI staining (red), (right panel).Adapted from Meng, Q. et al. (2015) Am. J. Physiol. 309, C1. with permission of The American Physiological Society.

    Last update: 25/11/2018

    Anti-ASIC3 Antibody (#ASC-018) is a highly specific antibody directed against an epitope of the rat protein. The antibody can be used in western blot, immunocytochemistry, and immunohistochemistry applications. It has been designed to recognize ASIC3 from rat, human, and mouse samples.

    For research purposes only, not for human use
    Citations
    Western blot citations
    1. Rat DRG lysate (1:200).
      Wu, Y. et al. (2017) Cell. Mol. Neurobiol. 37, 635.
    2. Mouse bone marrow derived macrophage (BMM) lysate.
      Kong, X. et al. (2013) Cell. Immunol. 281, 44.
    3. Human SH-SY5Y cell lysate.
      Xiong, Q.J. et al. (2012) Am. J. Physiol. 303, C376.
    4. Mouse immature dentritic cells.
      Tong, J. et al. (2011) J. Immunol. 186, 3686.
    Immunohistochemistry citations
    1. Rat DRG sections (1:50).
      Wu, Y. et al. (2017) Cell. Mol. Neurobiol. 37, 635.
    2. Rat trigeminal sections (1:50).
      Meng, Q. et al. (2015) Am. J. Physiol. 309, C1.
    3. Rat trigeminal ganglia (1:400).
      Yan, J. et al. (2013) Headache 53, 1250.
    4. Mouse anterial synovial sections (1:500).
      Kolker, S.J. et al. (2010) Ann. Rheum. Dis. 69, 903.
    Immunocytochemistry citations
    1. Mouse primary DRGs (1:100).
      Radu, B.M. et al. (2014) Cell Biochem. Biophys. 68, 9.
    2. Human SH-SY5Y cell lysate.
      Xiong, Q.J. et al. (2012) Am. J. Physiol. 303, C376.
    3. Mouse dendritic cells.
      Tong, J. et al. (2011) J. Immunol. 186, 3686.
    4. Mouse synoviocytes. Also tested in ASIC3-/- mice.
      Kolker, S.J. et al. (2010) Ann. Rheum. Dis. 69, 903.
    More product citations
    1. Ohbuchi, T. et al. (2010) J. Physiol. 588, 2147.
    2. Akiba, Y. et al. (2008) Gut 57, 1654.
    3. Ye, J.H. et al. (2007) Biochem. Biophys. Res. Commun. 355, 986.
    Related Products
      • Products for control experiments
        1. ASIC3 Channel Overexpressed Membrane Fractions (#LX-100). The ASIC3 overexpressed membrane fractions enables end users to check the expression of endogenous ASIC3 as opposed to a tagged version and serves as a positive control for testing the expression of the channel and a quick way to test the antibody.
        2. ASIC3 Channel Antibody and Membrane Fractions Kit (#LK-100). This kit includes the ASIC3 overexpressed membrane fractions, and Anti-ASIC3 antibody (#ASC-018).
      • Antibodies for immuno-colocalization experiments
        1. Guinea pig Anti-ASIC3 Antibody (#AGP-052) is raised in guinea pig and can be used in immuno-colocalization studies in conjunction with any of our antibodies raised in rabbit. This antibody has been raised against the same epitope as #ASC-018.