• Product name
  • Description
    Rabbit polyclonal to HCN4
  • Tested applications
    Suitable for: WB, ICC/IFmore details
    Unsuitable for: IHC-P
  • Species reactivity
    Reacts with: Mouse, Rat, Dog, Human
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1150 to the C-terminus of Human HCN4.

  • Positive control
    • WB: Human (fetal), mouse and rat heart tissue lysates. ICC/IF: PC12 cells.



Our Abpromise guarantee covers the use of ab69054 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB Use a concentration of 1 µg/ml. Detects a band of approximately 155 kDa (predicted molecular weight: 129 kDa).
ICC/IF Use a concentration of 10 µg/ml.
  • Application notes
    Is unsuitable for IHC-P.
  • Target

    • Relevance
      HCN4 is a member of the family of hyperpolarization activated and cyclic nucleotide gated (HCN) channels. HCN currents have been linked to pacemaker activity in the heart and brain, resting potential control, as well as neuronal plasticity. It has been shown that HCN4 channels function as receptors for sour taste, and are associated with pacemaker potential generation in the sinoatrial node.
    • Cellular localization
      Membrane; multi pass membrane protein.
    • Database links
    • Alternative names
      • HCN 4 antibody
      • Hyperpolarization activated cyclic nucleotide gated potassium channel 4 antibody
      • Potassium/sodium hyperpolarization activated cyclic nucleotide gated channel 4 antibody

    Anti-HCN4 antibody images

    • All lanes : Anti-HCN4 antibody (ab69054) at 1 µg/ml

      Lane 1 : Heart (Human) Whole Cell Lysate - fetal normal tissue
      Lane 2 : Heart (Rat) Tissue Lysate
      Lane 3 : Heart (Mouse) Tissue Lysate

      Lysates/proteins at 10 µg per lane.

      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
      Developed using the ECL technique

      Performed under reducing conditions.

      Predicted band size : 129 kDa
      Observed band size : 155 kDa (why is the actual band size different from the predicted?)
      Additional bands at : 55 kDa,70 kDa,78 kDa. We are unsure as to the identity of these extra bands.

      Exposure time : 90 seconds

      HCN4 contains a potential glycosylation site (SwissProt) which may explain its migration at a higher molecular weight than predicted. This higher band has been described in the following publications: PMID: 14657344, PMID: 19471099.

      This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab69054 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

    • Immunofluorescence image of HCN4 (ab69054) on human embryonic stem cell derived cardiomyocytes (in green). The cells were fixed in paraformaldehyde and permeabilized in 0.1% Triton-X-100.

      See Abreview

    • IHC-P image of HCN4 staining on mouse heart tissue sections using ab69054 (1:3000). The sections were deparaffinized and subjected to heat mediated antigen retrieval using citric acid. The sections were blocked using 1% BSA for 10 mins at 21°C. The sections were then incubaed with ab69054 (1:3000) for 2 hours at 21°C. The secondary antibody used Goat polyclonal to rabbit IgG (biotin) (1:250)

      See Abreview

    • ICC/IF image of ab69054 stained PC12 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab69054, 10µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 100% methanol fixed (5 min) PC12 cells at 10µg/ml.

    References for Anti-HCN4 antibody (ab69054)

    This product has been referenced in:
    • Feng Y  et al. Shox2 influences mesenchymal stem cell fate in a co-culture model in vitro. Mol Med Rep 14:637-42 (2016). WB ; Dog . Read more (PubMed: 27222368) »
    • He W  et al. Expression of hyperpolarization-activated cyclic nucleotide-gated channel isoforms in a canine model of atrial fibrillation. Exp Ther Med 12:433-436 (2016). WB . Read more (PubMed: 27347074) »

    See all 3 Publications for this product

    Product Wall

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Mouse Tissue sections (Heart)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citric acid
    Blocking step
    BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C

    Mr. Carl Hobbs

    Verified customer

    Submitted Feb 08 2013

    I am happy to hear that I was able to resolve this issue to your satisfaction. Please do not hesitate to contact me or anyone on the Scientific Support Team should you have any additional questions or require further assistance.

    Have a gr...

    Read More

    Thank you for your reply.

    I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement.

    To check the status of the order pl...

    Read More

    Thank you for contacting Abcam regarding ab69054.

    I am sorry that you have been experiencing difficulties with this antibody in WB and ICC. I have reviewed the protocol information and data you provided and I agree that this antibody is n...

    Read More
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Immunocytochemistry/ Immunofluorescence
    Human Cell (hESC-derived cardiomyocyte)
    hESC-derived cardiomyocyte
    Yes - 0.1% Triton X-100 in PBS
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 25°C

    Dr. David Anderson

    Verified customer

    Submitted Dec 03 2010

    Thank you for providing your protocol details. Upon initial review of your protocol, I have a few follow up questions and potential recommendations:

    1. You mentioned that your sections are 4% PFA fixed. Are they frozen or paraffin embedded fol...

    Read More


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