Anti-KV11.1 (HERG) (extracellular) Antibody

Voltage gated K+ channel subfamily H member 2, KCNH2, ether-a-go-go-related channel 1
    Cat #: APC-109
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: h, r
    May also work in: m
    Immunogen
    Peptide AFLLKETEEGPPATEC corresponding to residues 430-445 of human KV11.1 (HERG) (Accession Q12809).  Extracellular, between S1 and S2 domains.
    Accession (Uniprot) Number Q12809
    Gene ID 3757
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Rat, mouse - 11/16 amino acid residues identical; dog, rabbit - 15/16 amino 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 0.6 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 -20ºC.
    Preadsorption Control 2 μg peptide per 1 μg antibody.
    Standard quality control of each lot Western blot analysis.
    Applications: ic, ifc, ih, ip, lci, wb
    Western blot
    Western blot analysis of Kv11.1 transfected HEK-293 cells:
    1. Anti-KV11.1 (HERG) (extracellular) Antibody (#APC-109), (1:200).
    2. Anti-KV11.1 (HERG) (extracellular) Antibody, preincubated with the control peptide antigen.
    Immunoprecipitation
    Immunoprecipitation of KV11.1 expressing HEK-293 cells:
    1. Cell lysate.
    2. Lysate + protein A beads + Anti-KV11.1 (HERG) (extracellular) Antibody  (#APC-109).
    3. Lysate + protein A beads + Anti-KV11.1 (erg1) Antibody (#APC-016).
    4. Lysate + protein A beads + Anti-KV11.1 (HERG) Antibody (#APC-062).
    5. Lysate + protein A beads + pre-immune rabbit serum.
    Red arrow indicates KV11.1 while the black arrow shows the IgG heavy chain.
    Immunoblot was performed with Anti-KV11.1 (HERG) (extracellular) Antibody.
    Immunohistochemistry
    Expression of KV11.1 in rat heart
    Immunohistochemical staining of rat heart using Anti-KV11.1 (HERG) (extracellular) Antibody (#APC-109). A. Transversal section of the atrium wall; note that arterial smooth muscle fibers were not stained (green arrow). B. Longitudinal section  of the myocardium. C. Section showing myocardium and endocardium (red arrow). DAB product is brown and the counterstain is cresyl violet.
    Indirect flow cytometry
    Indirect flow cytometry analysis of human chronic myelogenous leukemia (K562) cells:
    ___ Unstained cells + goat-anti-rabbit-FITC.
    ___ Cells + Anti-KV11.1 (HERG) (extracellular) Antibody
    (#APC-109) + goat-anti-rabbit-FITC.
    The control antigen is not suitable for this application.
    Live cell imaging / Immunocytochemistry
    Expression of KV11.1 in HEK-293 transfected cells
    Immunocytochemical staining of live intact HEK-293 transfected cells.Cells were stained with Anti-KV11.1 (HERG) (extracellular) Antibody (#APC-109), (1:25) (left panel) or with KV11.1 (HERG) (extracellular) antibody preincubated with the control peptide antigen (right panel), followed by goat anti-rabbit-AlexaFluor-555 secondary antibody (red). Nuclei of the living cells were stained with the cell permeable dye Hoechst 33342 (blue). Note that not all cells are stained, indicating that not all the KV11.1 channels are expressed at the cell membrane at the moment of staining.
    References
    1. Curran, M.E. et al. (1995) Cell 80, 795.
    2. Sanguinetti, M.C. et al. (1995) Cell 81, 299.
    3. Pardo, L.A. et al. (2004) Physiology 19, 285.
    Scientific background

    The KV11.1 (HERG) channel is a member of the ether-a-go-go (EAG) subfamily of voltage-dependent K+channels that includes the related proteins KV11.2 and KV11.3 (erg2 and erg3). KV11.1 possess the signature structure of the voltage-dependent K+ channels: six membrane-spanning domains and intracellular N and C termini.

    The KV11.1 current is characterized by strong inward rectification with slow activation and very rapid inactivation kinetics. The channel is expressed in the brain and heart (where it underlies the IKr current) and has a central role in mediating repolarization of action potentials.

    Mutations in the KV11.1 channel cause inherited long QT syndrome (LQTS) or abnormalities in the repolarization of the heart that are associated with life-threatening arrhythmias and sudden death. All the identified KV11.1 mutations produce loss of function of the channel via several cellular mechanisms ranging from alterations of gating properties, alterations of channel permeability/selectivity and alterations in intracellular channel trafficking that decreases the number of channels that reach the cell membrane. Lately drug-induced forms of LQTS have been reported for a wide range of non-cardiac drugs including antihistamines, psychoactive agents and antimicrobials. All these drugs potently block the KV11.1 channel as an unintended side effect, prompting regulatory drug agencies to issue recommendations for the testing of new drugs for their potential KV11.1 blocking effect.

    In addition, KV11.1 expression was found to be upregulated in several tumor cell lines of different histogenesis suggesting that it confers the cells some advantage in cell proliferation. Indeed, in several studies it has been shown that inhibition of the KV11.1 current leads to a decrease in tumor cell proliferation.

    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: 31/07/2018

    Anti-KV11.1 (HERG) (extracellular) Antibody (#APC-109) is a highly specific antibody directed against an extracellular epitope of the human protein. The antibody can be used in western blot, immunoprecipitation, immunohistochemical, immunocytochemical and indirect flow cytometry applications. It has been designed to recognize KV11.1 from human, rat, and mouse samples.

    For research purposes only, not for human use
    Citations
    More product citations
    1. Crottes, D. et al. (2011) J. Biol. Chem. 286, 27947.
    2. Mihic, A. et al. (2011) PLoS One. 6, e18273.
    3. Smith, J.L. et al. (2011) Am. J. Physiol. Cell Physiol. 301, C75.
    4. Stump, M.P.  et al. (2011) Am. J. Physiol. Heart Circ. Physiol. 300, H312.
    5. Wu, Z-Y. et al. (2011) J. Biol. Chem. 286, 22186.
    6. Nof, E. et al. (2010) Circ. Cardiovasc. Genet 3, 199.
    7. Delisle, B.P. et al. (2009) J. Biol. Chem. 284, 2844.
    8. Potet, F. et al. (2009) J. Mol. Cell. Cardiol. 46, 257.
    9. Chtcheglova, L.A. et al. (2008) Pflugers Arch. 456, 247.
    10. Claassen, S. et al. (2008) J. Membr. Biol. 222, 31.
    11. Takemasa, H. et al. (2008) Br. J. Pharmacol. 153, 439.
    12. Wu, Z-Y. et al. (2008) Endocrinology 149, 5061.
    Related Products