Anti-CLC-3 (CLCN3) Antibody

Chloride channel 3, Chloride transporter ClC-3, H+/Cl- exchange transporter 3
    Cat #: ACL-001
    Alternative Name Chloride channel 3, Chloride transporter ClC-3, H+/Cl- exchange transporter 3
  • KO Validated
  • Lyophilized Powder
  • Antigen Incl.
  • Type: Polyclonal
    Host: Rabbit
    Reactivity: h, m, r
    • GST fusion protein with the sequence SLVVIVFELTGGLEYIVPLMAAVMTSKWVGDAFGREGIYEAHIRLNGYPFLDAKEEFTHTTLAADVMRPR, corresponding to amino acid residues 592-661 of rat CLC-3 (Accession P51792). Intracellular, near the C-terminus.
    Accession (Uniprot) Number P51792
    Gene ID 84360
    Peptide confirmation Confirmed by DNA sequence and SDS-PAGE.
    Homology Mouse - identical; human, rabbit, guinea pig - 69/70 amino acid residues identical; Xenopus laevis - 61/70 amino acid residues identical.
    Rat CLC-4 - 46/70 amino acid residues identical; rat CLC-5 - 49/70 amino acid residues identical.
    RRID AB_2039814.
    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, 5% sucrose, 0.025% 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).
    Negative 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.
    Negative control antigen reconstitution 100 μl PBS.
    Negative control antigen storage after reconstitution -20°C.
    Preadsorption Control 3 μg fusion protein per 1 μg antibody.
    Standard quality control of each lot Western blot analysis.
    Applications: ic, if, ih, ip, wb
    May also work in: ifc*
    Western blot
    • Anti-CLC-3 (CLCN3) Antibody
      Western blot analysis of rat brain membranes:
      1. Anti-CLC-3 (CLCN3) Antibody (#ACL-001), (1:200).
      Anti-CLC-3 (CLCN3) Antibody, preincubated with a control antigen.
    • Anti-CLC-3 (CLCN3) Antibody
      Western blot analysis of membranes from Xenopus oocytes, expressing CLC-3, CLC-4, and CLC-5, using Anti-CLC-3 (CLCN3) Antibody (#ACL-001) (kindly provided by Prof. Jordi Ehrenfeld, University of Nice).
    • Human glioma lysates (1:500), (Cuddapah, V.A. et al. (2013) J. Neurosci. 33, 1427.).
    • tsA cells expressing CLC-3 (Farmer, L.M. et al. (2013) J. Physiol. 591, 1001.).
    • Rat brain sections (1:100) (Farmer, L.M. et al. (2013) J. Physiol. 591, 1001.).
    • tsA cells expressing CLC-3 (Farmer, L.M. et al. (2013) J. Physiol. 591, 1001.).
    1. Jentsch, T.J. et al. (2002) Physiol. Rev. 82, 503.
    2. Babini, E. and Pusch, M. (2004) Physiology 19, 293.
    3. Stobrawa, S.M. et al. (2001) Neuron 29, 185.
    4. Moreland, J.G. et al. (2006) J. Biol. Chem. 281, 12277.
    Scientific background

    CLC-3 is a member of the voltage-dependent Cl- channel (CLC) family that includes nine known members in mammals. CLC channels can be classified as plasma membrane channels and intracellular organelle channels. The first group includes the CLC-1CLC-2 CLC-Ka and CLCKb channels. The second group comprises the CLC-3, CLC-4, CLC-5, CLC-6 and CLC-7.

    CLC channels that function in the plasma membrane are involved in the stabilization of membrane potential and in transepithelial transport. The presumed function of the intracellular CLC channels is support of the acidification of the intraorganellar compartment. In this regard, recent reports indicate that ClC-4 and ClC-5 (and by inference ClC-3) can function as Cl-/H+ antiporters.1, 2

    The functional unit of the CLC channels is a dimer with each subunit forming a proper pore. Although the crystal structure of bacterial CLC channels was resolved, the topology of the CLC channels is complex and has not been fully elucidated. It is generally accepted that both the N- and C- terminus domains are intracellular while the number and configuration of the transmembrane domains vary greatly between different models. 1,2

    CLC-3 is widely distributed with prominent expression in tissues of neuroectoderm origin. In the brain, it is highly expressed in the hippocampus, olfactory bulb and olfactory cortex. The channel is also prominently expressed in aortic and coronary vascular smooth muscle cells, aortic endothelial cells and tracheal and alveolar epithelial cells.

    The physiological function of CLC-3 is not entirely clear, but it has been suggested that CLC-3 generates a shunt current of chloride for v-H+-ATPases, thereby aiding the acidification of endosomes and synaptic vesicles as well as lysosomes. Disruption of the ClC-3 gene in mice causes severe neuronal loss, leading to a complete loss of the hippocampus in adult mice. In addition, CLC-3 has been shown to have a critical role in the respiratory burst and phagocytosis of polymorphonuclear cells, a key cell type of innate host defense. 3,4

    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:

    Anti-CLC-3 (CLCN3) Antibody
    Colocalization and expression of KCa3.1, ClC-3 and B2R in glioma cells.A-C. Immunocytochemical staining of human D54 glioma cells using Mouse Anti-KCNN4 (KCa3.1, SK4) (extracellular) Antibody (#ALM-051). KCNN4, ClC-3 and B2R co-localize to the edges of cells. D. Western blot analysis of a number of glioma cells using Anti-CLC-3 (CLCN3) Antibody (#ACL-001) and Mouse Anti-KCNN4 (KCa3.1, SK4) (extracellular) Antibody.Adapted from Cuddapah, V.A. et al. (2013) with permission of the Society for Neuroscience.

    Last update: 24/01/2020

    Anti-CLC-3 (CLCN3) Antibody (#ACL-001) is a highly specific antibody directed against an epitope of the rat protein. The antibody can be used in western blot and immunohistochemistry applications. It has been designed to recognize CLC-3 from rat, mouse, and human samples.

    For research purposes only, not for human use



    Scientific Background


    Published figures using this product
    • Anti-CLC-3 (CLCN3) Antibody

      Expression of CLC-3 in human D54 glioma cells.

      Immunocytochemical staining of human D4 glioma cells using Anti-CLC-3 (CLCN3) Antibody (#ACL-001), (green). CLC-3 co-localizes with cortical actin (stained with phalloidin, red) on the plasma membrane. DAPI (blue) is used to stain nuclei.
      Adapted from Cuddapah, V.A. and Sontheimer, H. (2010) J. Biol. Chem. 285, 2010. with permission of the American Society for Biochemistry and Molecular Biology.
    KO validation citations
    1. Immunohistochemical staining and western blot analysis of mouse DRG sections and lysates respectively (1:200). Tested in CLC3-/- mice.
      Pang, R.P. et al. (2016) Neuropharmacology 110, 181.
    Western blot citations
    1. Mouse DRG lysate (1:500).
      Pang, R.P. et al. (2016) Neuropharmacology 110, 181.
    2. Human fibroblast lysate.
      Lakoma, J. et al. (2016) J. Cell. Physiol. 231, 192.
    3. Human mG-63 osteosarcoma cell lysate.
      Du, S. and Yang, L. (2015) Int. Clin. Exp. Pathol. 8, 1622.
    4. Mouse endothelial progenitor cells (1:500).
      Liu, J. et al. (2013) Apoptosis 18, 1262.
    5. tsA cells expressing CLC-3.
      Farmer, L.M. et al. (2013) J. Physiol. 591, 1001.
    6. Human glioma lysates (1:500).
      Cuddapah, V.A. et al. (2013) J. Neurosci. 33, 1427.
    Immunoprecipitation citations
    1. tsA cells expressing CLC-3.
      Farmer, L.M. et al. (2013) J. Physiol. 591, 1001.
    Immunohistochemistry citations
    1. Mouse DRG sections (1:200). Also tested in CLC3-/- mice.
      Pang, R.P. et al. (2016) Neuropharmacology 110, 181.
    2. Rat brain sections (1:100).
      Farmer, L.M. et al. (2013) J. Physiol. 591, 1001.
    Immunocytochemistry citations
    1. sA cells expressing CLC-3.
      Farmer, L.M. et al. (2013) J. Physiol. 591, 1001.
    2. Human D54 glioma cells.
      Cuddapah, V.A. and Sontheimer, H. (2010) J. Biol. Chem. 285, 2010.
    More product citations
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    2. Cuddapah, V.A. et al. (2012) Am. J. Physiol. 302, C527.
    3. Ohtaki, H. et al. (2012) J. Neurosci. Res. 90, 2163.
    4. Wang, L. et al. (2012) Am. J. Physiol. 303, C14.
    5. Yang, L. et al. (2011) J. Cell. Physiol. 226, 2516.
    6. Liu, Y.J. et al. (2010) Hypertension 56, 445.
    7. Schenck, S. et al. (2009) Nat. Neurosci. 12, 156.
    8. Habela, C.W. et al. (2008) J. Neurosci. 28, 9205.
    9. Li, H. et al. (2008) Arch Otolaryngol Head Neck Surg. 134, 301.
    10. Yin, Z. et al. (2008) Am. J. Physiol. Cell Physiol. 294, C535.
    11. Kasinathan, R.S. et al. (2007) FEBS Lett. 581, 5407.
    12. Li, S.J. et al. (2006) J. Biochem. 139, 813.
    13. Lemonnier, L. et al. (2005) Endocr. Relat. Cancer 12, 335.
    14. Hara-Chikuma, M. et al. (2005) J. Biol. Chem. 280, 1241.
    15. Zhou, J-G. et al. (2005) J. Biol. Chem. 280, 7301.
    16. Fadool, D.A. et al. (2004) Neuron 41, 389.
    17. Lemonnier, L. et al. (2004) Cancer Res. 64, 4841.
    18. Robinson, N.C. et al. (2004) J. Physiol. 556.2, 353.
    19. Koni, P.A. et al. (2003) J. Biol. Chem. 278, 39443.
    20. Olsen, M.L. et al. (2003) J. Neurosci. 23, 5572.
    21. Hermoso, M. et al. (2002) J. Biol. Chem. 277, 40066.
    22. Tsuboi, T. et al. (2002) Biophys. J. 83, 172.
    23. Weng, T.X. et al. (2002) Am. J. Physiol. Cell Physiol. 283, 839.
    24. Duan, D. et al. (2001) J. Physiol. 531, 437.
    25. Britton, F.C. et al. (2000) Am. J. Physiol. 279, H2225.
    26. Weylandt, K-H. et al. (2001) J. Biol. Chem. 276, 17461.
    27. Wang, L. et al. (2000) J. Physiol. 524.1, 63.
    28. Wills, N.K. et al. (2000) Invest. Ophalmol. Vis. Sci. 41, 4247.
    29. Shimada, K. et al. (2000) Am J. Physiol. 279, G268.
    30. Luyckx, V.A. et al. (1999) Proc. Natl. Acad. Sci. USA 96, 12174.
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