Anti-HCN4 Antibody

Hyperpolarization-activated cyclic nucleotide-gated potassium channel 4
    Cat #: APC-052
    Alternative Name Hyperpolarization-activated cyclic nucleotide-gated potassium channel 4
  • KO Validated
  • Lyophilized Powder
  • Antigen Incl.
  • Type: Polyclonal
    Host: Rabbit
    Reactivity: h, m, r
      • GST fusion protein with the sequence HGHLHDSAEERRLIAEGDASPG EDRTPPGLAAEPERP, corresponding to amino acid residues 119-155 of human HCN4 (Accession Q9Y3Q4). Intracellular, N-terminus.
        Anti-HCN4 Antibody
    Accession (Uniprot) Number Q9Y3Q4
    Gene ID 59266
    Peptide confirmation Confirmed by DNA sequence and SDS-PAGE.
    Homology Rabbit - identical; rat - 35/37 amino acid residues identical.
    RRID AB_2039906.
    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).
    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*
      • Anti-HCN4 Antibody
        Western blot analysis of rat brain membranes:
        1. Anti-HCN4 Antibody (#APC-052), (1:200).
        2. Anti-HCN4 Antibody, preincubated with the control peptide antigen.
        Human HCN4 transfected in HEK-293T cells. (Nof, E. et al. (2007) Circulation 116, 463.).
      • Rat thalamus lysates (Whitaker, G.M. et al. (2007) J. Biol. Chem282, 22900.).
      • Anti-HCN4 Antibody
        Expression of HCN4 in mouse thalamus
        Immunohistochemical staining of mouse thalamus using Anti-HCN4 Antibody (#APC-052). A. HCN4 (red) appears in the neuropil of the lateral nucleus (LN). B. Staining of reticular nucleus (RN) with mouse anti-Parvalbumin. C. Confocal merge of HCN4 and Parvalbumin images demonstrates separate localization of these proteins in the thalamus.
      • Mouse sinoatrial myocytes (1:200) (St. Clair, J.R. et al. (2015) Am. J. Physiol. 309, H490).
    References
      • Hyperpolarization-activated cation currents (Ih) appear in the heart and the brain and have 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 is comprised of four homologous members, HCN1-4.

        Each HCN subunit consists of six transmembrane domains (TM), a pore region between TM5-TM6 and a binding domain for cyclic nucleotides (CNBD) in the cytoplasmic C-terminus.2

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

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

        HCN4 is highly expressed in a restricted manner in adult sinoatrial (SA) node, constituting a good molecular marker for the adult cardiac pacemaker and might serve as a unique marker of the developing SA node .4,5

        mRNA expression of HCN4 is most abundant in medial habenula and anterior and principal relay nuclei of the thalamus.6

    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-HCN4 Antibody

    Knockout validation of Anti-HCN4 Antibody in mouse heart.Western blot analysis of mouse heart lysate using Anti-HCN4 Antibody (#APC-052). HCN4 immunodetection in wild-type, overexpressed, and knockdown animals. HCN4 expression is abolished in knockdown animals. In addition, kidney lysates were loaded as a negative control, a tissue which does not express HCN4. α-tubulin is a loading control.Adapted from Kozasa, Y. et al. (2018) J. Physiol. 596, 809. with permission of The Physiological Society.

    Last update: 08/11/2019

    Anti-HCN4 Antibody (#APC-052) is a highly specific antibody directed against an epitope of the human protein. The antibody can be used in western blot, immunoprecipitation, immunocytochemistry, and immunohistochemistry applications. It has been designed to recognize HCN4 from human, rat, and mouse samples.

    For research purposes only, not for human use
    Citations
      • Anti-HCN4 Antibody
        Expression of HCN4 in mouse sinoatrial myocytes.
        Immunocytochemical staining of mouse cultured (right panel) and acutely isolated (left panel) sinoatrial myocytes using Anti-HCN4 Antibody (#APC-052).
        Adapted from St. Clair, J.R. et al. (2015) Am. J. Physiol. 309, H490. with permission of the American Physiological Society.
      • Western blot analysis of mouse heart lysates. Tested in HCN4 knockdown mice.
        Kozasa, Y. et al. (2018) J. Physiol. 596, 809.
      • Mouse heart lysates. Also tested in HCN4 knockdown mice.
        Kozasa, Y. et al. (2018) J. Physiol. 596, 809.
      • Rat heart lysate (1:200).
        Atkinson, A.J. et al. (2013) J. Am. Heart Assoc. 2, e000246.
      • Human HCN4 transfected in HEK-293T cells.
        Nof, E. et al. (2007) Circulation 116, 463.
      • Rat thalamus lysates.
        Whitaker, G.M. et al. (2007) J. Biol. Chem. 282, 22900.
      • Mouse heart sections.
        Samal, E. et al. (2019) Front. Physiol. 10, 235.
      • Mouse heart sections.
        Bhattacharyya, S. et al. (2017) PLoS ONE 12, e0174517.
      • Mouse embryo sections.
        Fujii, M. et al. (2017) Nat. Commun. 8, 14664.
      • Mouse retina sections (1:500).
        Puller, C. et al. (2017) PLoS ONE 12, e0173455.
      • Rat diaphragm sections (1:25).
        Negrini, D. et al. (2016) Am. J. Physiol. 311, H892.
      • Mouse heart sections.
        Harris, J.P. et al. (2015) Mol. Cell. Biol. 35, 649.
      • Rat heart sections.
        Mackiewicz, U. et al. (2014) J. Cell. Physiol. 229, 813.
      • Mouse heart sections.
        Mahida, S. et al. (2014) Cardiovasc. Res. 101, 326.
      • Rat heart sections (1:20).
        Atkinson, A.J. et al. (2013) J. Am. Heart Assoc. 2, e000246.
      • Mouse eye sections (1:500).
        Zhang, J. et al. (2013) Synapse 67, 525.
      • Mouse sinoatrial myocytes (1:200).
        St. Clair, J.R. et al. (2015) Am. J. Physiol. 309, H490.
      • HEK-293 transfected cells (1:200).
        Altomare, C. et al. (2003) J. Physiol. 549, 347.
      • Lee, M.Y. et al. (2017) PLoS ONE 12, e0176031.
      • Semmler, J. et al. (2014) Cell. Physiol. Biochem. 34, 1199.
      • Hoffmann, S. et al. (2013) Basic Res. Cardiol. 108, 339.
      • Nakashima, N. et al. (2013) J. Physiol. 591, 1749.
      • Weisbrod, D. et al. (2013) Proc. Natl. Acad. Sci. U.S.A. 110, E1685.
      • Frank, M. et al. (2012) Circ. Res. 111, 1528.
      • Doan, T.N. et al. (2004) J. Neurosci. 24, 3335.
      • Much, B. et al. (2003) J. Biol. Chem. 278, 43781.
      • Han, W. et al. (2002) Circ. Res. 91, 790.
    Related Products
        • Guinea pig Anti-HCN4 Antibody (#AGP-004) is raised in guinea pig and can be used in immuno-colocalization studies in conjunction with any of our antibodies raised in rabbit. This antibody has been raised against the same epitope as #APC-052.