Anti-CLC-2 (CLCN2) Antibody

Chloride channel 2, ClC2
Cat #: ACL-002
Alternative Name Chloride channel 2, ClC2
  • KO Validated
  • Lyophilized Powder
  • Antigen Incl.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: h, m, r
      • Peptide RSRHGLPREGTPSDSDDKC, corresponding to amino acid residues 888-906 of rat CLC-2. (Accession P35525). Intracellular, C-terminus.
    Accession (Uniprot) Number P35525
    Gene ID 29232
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Mouse - identical; rabbit, swine - 18/19 amino acid residues identical; human - 17/19 amino acid residues identical; guinea pig - 15/19 amino acid residues identical.
    Purity Affinity 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 50 μl or 0.2 ml double distilled water (DDW), depending on the sample size.
    Antibody concentration after reconstitution 0.6 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 analysis of rat brain membranes:
        1. Anti-CLC-2 (CLCN2) Antibody (#ACL-002), (1:200).
        2. Anti-CLC-2 (CLCN2) Antibody
        , preincubated with the control peptide antigen.
      • Rat brain membranes.
      • Human fibroblasts (1:100) (Lakoma, J. et al. (2016) J. Cell. Physiol. 231, 192.).
      • CLC-2 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-1, CLC-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-2 is widely distributed with prominent expression in brain, kidney, lung and the gastrointestinal system.  Mutations in the CLC-2 channel gene detected in humans are associated with idiopathic generalized epilepsies while disruption of the ClC-2 gene in mice is associated with testicular and retinal degeneration. 3,4

    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-CLC-2 (CLCN2) Antibody
    Knockout validation of Anti-CLC-2 (CLCN2) Antibody in parietal cells of mouse gastric mucosa.
    A. Immunohistochemical staining of mouse gastric mucosa using Anti-CLC-2 (CLCN2) Antibody (#ACL-002) (red). Same sections are stained for H+/K+ ATPase (green). Merge panel shows co-localization of the two proteins. B. Western blot analysis of ClC-2 in WT and ClC-2-/- mouse gastric mucosa using Anti-CLC-2 (CLCN2) Antibody shows that antibody is indeed specific for ClC-2. C. Immunohistochemical staining of mouse gastric mucosa shows lack of ClC-2 staining in ClC-2-/- mouse. D. Immunogold electronmicroscopy of ClC-2 in WT and ClC-2-/- mouse gastric parietal cells.Adapted from Nighot, M.P. et al. (2015) PLoS ONE 10, e0138174. with kind permission of Dr. Blikslager, A.T. North Carolina State University, College of Veterinary Medicine, Raleigh, North Carolina, United States of America.
    Last update: 04/06/2019

    Anti-CLC-2 (CLCN2) Antibody (#ACL-002) is a highly specific antibody directed against an epitope of the rat protein. The antibody can be used in western blot, immunohistochemistry and immunocytochemistry applications. It has been designed to recognize CLC-2 channel from rat, mouse, and human samples.

    For research purposes only, not for human use
    Citations
      • Western blot analysis of mouse gastric mucosa lysate. Immunohistochemical staining of mouse gastric tissue sections. Also tested in ClC-2-/- mouse gastric parietal cells.
        Nighot, M.P. et al. (2015) PLoS ONE 10, e0138174.
      • Human fibroblasts (1:200).
        Lakoma, J. et al. (2016) J. Cell. Physiol. 231, 192.
      • Western blot analysis of mouse gastric mucosa lysate. Also tested in ClC-2-/- mouse gastric parietal cells.
        Nighot, M.P. et al. (2015) PLoS ONE 10, e0138174.
      • Mouse gastric tissue sections. Also tested in ClC-2-/- mouse gastric parietal cells.
        Nighot, M.P. et al. (2015) PLoS ONE 10, e0138174.
      • Mouse nasal cavity sections.
        Schiffhauer, E.S. et al. (2013) Am. J. Physiol. 304, L324.
      • Human fibroblasts (1:100).
        Lakoma, J. et al. (2016) J. Cell. Physiol. 231, 192.
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