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Anti-Aquaporin 4 (AQP4) (249-323) Antibody

WCH4, Mercurial-insensitive water channel

Cat #: AQP-004
Alternative Name WCH4, Mercurial-insensitive water channel
  • KO Validated
  • Lyophilized Powder yes
    Type: Polyclonal
    Host: Rabbit
    Reactivity: h, m, r
    Immunogen
    • GST fusion protein with the sequence EYVFCPDVELKRRLKEAFSKAAQQTKGSYMEVEDNRSQVETEDLILKPGVVHVIDIDRGDEKKGKDSSGEVLSSV, corresponding to amino acid residues 249-323 of rat AQP4 (Accession P47863). Intracellular, C-terminus.
    Accession (Uniprot) Number P47863
    Gene ID 25293
    Peptide confirmation Confirmed by DNA sequence and SDS-PAGE.
    Homology Mouse - 73/75 amino acid residues identical; bovine - 71/75 amino acid residues identical; human - 69/75 amino acid residues identical; rabbit - 64/75 amino acid residues identical.
    RRID AB_2039734.
    Purity The serum was depleted of anti-GST antibodies by affinity chromatography on immobilized GST and then the IgG fraction was 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.
    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).
    Standard quality control of each lot Western blot analysis.
    Applications: ic, if, ifc, ih, wb
    May also work in: ip*
    Western blot
    • Western blot analysis of rat brain membranes:
      Western blot analysis of rat brain membranes:
      1. Anti-Aquaporin 4 (AQP4) (249-323) Antibody (#AQP-004), (1:1000).
      2. Anti-Aquaporin 4 (AQP4) (249-323) Antibody, preincubated with the control fusion protein (BLP-QP004).
    Immunohistochemistry
    • Rat brain (1:200) (Jo, S.Met al. (2011) Neurosci. Lett. 501, 25.). 

      Mouse kidney (Langaa, S. et al. (2012) Am. J. Physiol. 302, F1034.).
    Immunocytochemistry
    • Human AQP4 transfected in HEK-293 cells. (De Vidi, I. et al. (2011) Clin. Immunol. 138, 239.).
    Indirect flow cytometry
    • Human AQP4 transfected in HEK-293 cells. (De Vidi, I. et al. (2011) Clin. Immunol. 138, 239.).
    • The blocking peptide is not suitable for this application.
    References
    1. King, L.S. et al. (2004) Nat. Rev. Mol. Cell Biol. 5, 687.
    2. Manley, G.T. et al. (2004) Neuroscience. 129, 983.
    3. Lennon, V.A. et al. (2005) J. Exp. Med. 202, 473.
    Scientific background

    Aquaporin 4 (AQP-4) belongs to a family of membrane proteins that allow passage of water and certain solutes through biological membranes. The family is composed of 13 members (AQP-0 to AQP-12).

    The aquaporins can be divided into two functional groups based on their permeability characteristics: the aquaporins that are only permeated by water and the aquaglyceroporins that are permeated by water and other small solutes such as glycerol. AQP-4 together with AQP-1, AQP-2 and AQP-5 belong to the first group1. Little is known about the function of the two newest members, AQP-11 and AQP-12.

    The proteins present a conserved structure of six transmembrane domains with intracellular N- and C-termini. The functional channel is a tetramer but each subunit has a separate pore and therefore the functional channel unit, contains four pores1.

    AQP-4 is the major membrane water channel in the central nervous system. The channel is expressed in astrocyte foot processes in direct contact with capillary vessels in the brain suggesting a role in water transport under normal and pathological conditions. Indeed, transgenic mice lacking AQP-4 have reduced brain swelling and improved neurological outcome following water intoxication and focal cerebral ischemia. In contrast, brain swelling and clinical outcome are worse in AQP-4-null mice in models of vasogenic (fluid leak) edema caused by freeze-injury and brain tumor, probably due to impaired AQP-4-dependent brain water clearance2.

    In addition, it has been recently shown that neuromyelitis optica (NMO), an inflammatory demyelinating disease that selectively affects optic nerves and spinal cord, is caused by the development of an autoantibody directed against AQP-43.

    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
    Image & Title:

    Knockout validation of Anti-Aquaporin 4 (AQP4) (249-323) Antibody in mouse retina.Immunohistochemical staining of mouse retina sections using Anti-Kir4.1 (KCNJ10) Antibody (#APC-035) and Anti-Aquaporin 4 (AQP4) (249-323) Antibody (#AQP-004). Kir4.1 staining (green) and AQP4 immunoreactivity (red) are polarized with greatest expression in inner retina containing the end foot processes. Lack of AQP4 staining in AQP4-/- mice (lower panels). DAPI (blue) is used to stain nuclei.Adaped from Ruiz-Ederra, Jet al. (2007) J. Biol. Chem. 282, 21866. with permission of the American Society for Biochemistry and Molecular Biology.
    Last update: 11/04/2021

    Alomone Labs is pleased to offer a highly specific antibody directed against an epitope located at the intracellular C-terminal domain of rat AQP4. Anti-Aquaporin 4 (AQP4) (249-323) Antibody (#AQP-004) can be used in western blot, immunocytochemical, immunohistochemical and indirect flow cytometry applications. It recognizes the Aquaporin 4 channel from rat, mouse and human samples.

    For research purposes only, not for human use

    Applications

    Specifications

    Scientific Background

    Citations

    Citations
    Published figures using this product
    • AQP4 Serine phosphorylation is not required for plasma membrane trafficking.
      AQP4 Serine phosphorylation is not required for plasma membrane trafficking.
      Immunocytochemical staining of C6 rat glial cells transfected with wild type and AQP4 mutants. Cells were stained with Anti-Aquaporin 4 (AQP4) (249-323) Antibody (#AQP-004). Compared to wild type, none of the Serine-to-Alanine AQP4 mutants are affected by their ability to reach the plasma membrane.
      Adapted from Assentoft, M. et al. (2014) with permission of American Physiological Society.
    • Expression of Aquaporin 4 in mouse brain.
      Expression of Aquaporin 4 in mouse brain.
      Immunohistochemical staining of mouse brain sections using Anti-Aquaporin 4 (AQP4) (249-323) Antibody (#AQP-004). AQP4 staining (green) is detected in the cortex and hippocampus of normal mouse brain (NTG).
      Adapted from Zhang, X. et al. (2017) Front. Neurosci. 11, 212. with permission of Frontiers.
    KO validation citations
    1. Western blot analysis and immunohistochemical staining of mouse retina. Tested in AQP4-/- mice. 
      Ruiz-Ederra, J. et al. (2007) J. Biol. Chem. 282, 21866.
    Western blot citations
    1. Mouse primary astroglia cell lysate.
      Rocchio, F. et al. (2019) Cell Death Dis. 10, 24.
    2. Mouse retina lysate. Also tested in AQP4-/- mice. 
      Ruiz-Ederra, J. et al. (2007) J. Biol. Chem. 282, 21866.
    Immunohistochemistry citations
    1. Mouse brain and spinal cord sections (1:200).
      Guo, Y. et al. (2016) Mol. Neurobiol. 53, 3235.
    2. Human thyroid sections.
      Soelberg, K. et al. (2016) Neurol. Neuroimmunol. Neuroinflamm. 3, e252.
    3. Mouse kidney.
      Langaa, S. et al. (2012) Am. J. Physiol. 302, F1034.
    4. Rat brain (1:200).
      Jo, S.M. et al. (2011) Neurosci. Lett. 501, 25.
    5. Immunohistochemistry of mouse retina sections. Also tested in AQP4-/- mice. 
      Ruiz-Ederra, J. et al. (2007) J. Biol. Chem. 282, 21866.
    Immunocytochemistry citations
    1. Mouse primary astroglial cultures.
      Rocchio, F. et al. (2019) Cell Death Dis. 10, 24.
    2. Mouse muller cells.
      Vacca, O. et al. (2016) Hum. Mol. Genet. 25, 3070.
    3. C6 rat glia transfected cells.
      Assentoft, M. et al. (2014) Am. J. Physiol. 307, C957.
    4. Madin-Darby canine kidney cells (MDCK) (1:3000).
      Arnspang, E.C. et al. (2013) PLoS ONE 8, e73977.
    5. Xenopus oocytes (1:5000).
      Assentoft, M. et al. (2013) Glia 61, 1101.
    6. Human AQP4 transfected in HEK-293 cells.
      De Vidi, I. et al. (2011) Clin. Immunol. 138, 239.
    Indirect flow cytometry citations
    1. Human AQP4 transfected in HEK-293 cells.
      De Vidi, I. et al. (2011) Clin. Immunol. 138, 239.
    More product citations
    1. Toft-Bertelsen, T.L. et al. (2017) J. Physiol. 595, 3287.
    2. Zhang, X. et al. (2017) Front. Neurosci. 11, 212.
    3. Tham, D.K. et al. (2016) PLoS ONE 11, e0165439.
    4. Yde, J. et al. (2016) Front. Nutr. 3, 46.
    5. Kortenoeven, M.L.A. et al. (2013) J. Physiol. 591, 2205.
    6. Lien, C.F. et al. (2012) J. Biol. Chem. 287, 41374.
    7. Tham, D.K. and Moukhles, H. (2011) Am. J. Physiol. Renal Physiol. 301, F396.
    8. Merigo, F. et al. (2011) Dev. Neurobiol. 71, 854.
    9. Ikarashi, N, et al. (2011) Biol. Pharm. Bull. 34, 238.
    10. Noël, G. et al. (2010) PLoS One. 6, e17559.
    11. Satake, M. et al. (2010) Biol. Pharm. Bull. 33, 1965.
    12. Skjolding, A.D, et al. (2010) Cereb. Fluid Res. 7, 20.
    13. Toft-Hansen, H. et al. (2010) Glia 59, 166.
    14. Wang, Y-F. et al. (2009) J. Neurosci. 29, 1743.
    15. Zhang, H. and Verkman, A.S. (2008) Mol. Cell. Neurosci. 37, 1.
    16. Hibino, H. et al. (2007) Eur. J. Neurosci. 26, 2539.
    17. Ruiz-Ederra, J. et al. (2007) J. Biol. Chem. 282, 21866.
    18. Hibino, H. et al. (2004) J. Biol. Chem. 279, 44065.
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