Anti-HCN3 (extracellular) Antibody

K+/Na+ Hyperpolarization-activated cyclic nucleotide-gated channel 3
    Cat #: APC-083
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: h, m, r
    Immunogen
    Peptide (C)ELEPRLDAEVYK, corresponding to amino acid residues 190-201 of rat HCN3 (Accession Q9JKA8). 2nd extracellular loop.
    Accession (Uniprot) Number Q9JKA8
    Gene ID 114245
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Mouse, human – 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 0.85 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, lci, wb
    May also work in: ifc, ih, ip
    Western blot
    Western blot analysis of rat brain (lanes 1 and 3) and mouse brain (lanes 2 and 4) lysates:
    1,2. Anti-HCN3 (extracellular) Antibody (#APC-083), (1:200).
    3, 4. Anti-HCN3 (extracellular) Antibody preincubated with the control peptide antigen.
     
    Note: Samples were heated to 37°C prior to loading on gel.
    Live cell imaging / Immunocytochemistry
    Expression of HCN3 in rat U-87 MG cells
    Immunocytochemical staining of intact living U-87 MG cells. A. Extracellular staining of cells using Anti-HCN3 (extracellular) Antibody (#APC-083), (1:25) followed by goat anti-rabbit-AlexaFluor-594 secondary antibody (red). B. Live view of the cells. C. Extracellular staining merged with live view of the cells.
    References
    1. Much, Bet al (2003) J.Biol.Chem278, 43781.
    2. Notomi, T. and Shigemoto R. (2004) J. Comp. Neurol471, 241.
    Scientific background

    Hyperpolarization-activated cation currents (Ih) appear in the heart and the brain and play a crucial role in controlling electrical pacemaker activity, contributing to biological processes such as heartbeat, sleep-wake cycle and synaptic plasticity.1,2

    The Ih currents are generated by the Hyperpolarization-activated Cyclic Nucleotide-gated channel family (HCN), which comprises of four homologous members, named HCN1-4. Each HCN subunit consist of six transmembrane domains (TM), a pore region between TM5-TM6 and a binding domain to cyclic nucleotides (CNBD) in the cytoplasmic C-terminus.2

    The HCN subunits can form functional homomers and can also co-assemble into functionally heteromers.2

    The channels are closely related to each other and share homology of about 60%. However, they differ from each other in their cytoplasmic N- and the C-terminus. HCN1-4 mainly differ from each other with regard to their speed of activation and the extent by which they are modulated by cAMP. HCN1 is the fastest channel, followed by HCN2HCN3 and HCN4.2,3

    The HCN3 channels are abundant in the brain where high expression of HCN3 mRNA was found in the hypothalamus.

    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: 29/10/2018

    Alomone Labs is pleased to offer a highly specific antibody directed against an epitope of rat HCN3. Anti-HCN3 (extracellular) Antibody (#APC-083) can be used western blot and live cell imaging applications. The antibody recognizes an extracellular epitope and is thus ideal for detecting the channel in living cells. It has been designed to recognize HCN3 from human, rat and mouse samples.

    For research purposes only, not for human use
    Citations
    Related Products
      • Products for control experiments
        1. Anti-HCN3 Antibody (#APC-057).
          This antibody recognizes a different epitope and thus can also be used as a control for detecting HCN3.