Guinea pig Anti-HCN4 Antibody

Hyperpolarization-activated cyclic nucleotide-gated potassium channel 4
    Cat #: AGP-004
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Guinea pig
    Reactivity: h, m, r
    Immunogen
    GST fusion protein with sequence HGHLH DSAEE RRLIA EGDAS PGEDR TPPGL AAEPE RP, corresponding to amino acid residues 119-155 of human HCN4 (Accession Q9Y3Q4). Intracellular, N-terminus.
    Accession (Uniprot) Number Q9Y3Q4
    Gene ID 59266
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Rabbit - identical; rat - 35/37 amino acid residues identical; mouse - 33/37 amino acid 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 0.8 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).
    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: ih, wb
    May also work in: ic, ifc, ip
    Western blot
    Western blot analysis of rat brain lysate:
    1. Guinea pig Anti-HCN4 Antibody (#AGP-004), (1:200).
    2. Guinea pig Anti-HCN4 Antibody, preincubated with the control antigen.
    Immunohistochemistry
    Expression of HCN4 in mouse thalamus
    Immunohistochemical staining of mouse thalamus using Guinea pig Anti HCN4 Antibody (#AGP-004). A. HCN4 staining (green) appears in the neuropil of the ventral posterior thalamic nucleus (VPL). B. Nuclear staining using DAPI as the counter stain (blue). C. Merged images of A and B.
    Immuno-colocalization of HCN4 and HCN2 in mouse thalamus
    Immunohistochemical staining of mouse thalamus frozen section using Guinea pig Anti-HCN4 Antibody (#AGP-004) and rabbit Anti-HCN2 Antibody (#APC-030). A. Staining of HCN4 (green) appears in the ventral posterior thalamic nucleus (VPL). B. In the same section as in A, staining of HCN2 (red) appears in the ventral posterior thalamic nucleus (VPL) and also in the reticular thalamic nucleus (RT). The area between these thalamic nuclei (star) is white matter and neither protein is expressed in that region. C. Merged images of A and B.
    References
    1. Much, B. et al (2003) J.Biol.Chem278, 43781.
    2. Notomi, T. et al (2004) J. Comp. Neurol471, 241.
    3. Gravante, B. et al (2004) J.Biol.Chem279, 43847.
    4. Moosmang, S. et al. (2001) Eur. J. Biochem. 268, 1646.
    5. Garcia-Frigola, C. et al. (2003) Gene Expr Patterns 3, 777.
    6. Monteggia, L. et al. (2000) Molec. Brain Res. 1-2, 129.
    Scientific background

    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 HCN2, HCN3 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, 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: 30/10/2018

    Guinea pig Anti-HCN4 Antibody (#AGP-004), raised in guinea pigs, is a highly specific antibody directed against an epitope of the human protein. The antibody can be used in western blot and immunohistochemistry applications. It has been designed to recognize HCN4 from human, rat and mouse samples. The antigen used to immunize guinea pigs is the same as Anti-HCN4 Antibody (#APC-052) raised in rabbit. Our line of guinea pig antibodies enables more flexibility with our products such as immuno-colocalization studies, immunoprecipitation, etc.

    For research purposes only, not for human use