製品の概要

  • 製品名Anti-HCN4 antibody
    HCN4 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to HCN4
  • アプリケーション適用あり: WB, ICC/IF, IHC-Pmore details
  • 種交差性
    交差種: Mouse, Rat, Human
  • 免疫原

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

  • ポジティブ・コントロール
    • WB: Human (fetal), mouse and rat heart tissue lysates. ICC/IF: PC12 cells.

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
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.
IHC-P 1/3000.

ターゲット情報

  • 関連性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.
  • 細胞内局在Membrane; multi pass membrane protein.
  • 参照データベース
  • 別名
    • 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 画像

  • 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.

    Secondary
    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.

Anti-HCN4 antibody (ab69054) 使用論文

This product has been referenced in:
  • Kretschmannova K  et al. The expression and role of hyperpolarization-activated and cyclic nucleotide-gated channels in endocrine anterior pituitary cells. Mol Endocrinol 26:153-64 (2012). WB ; Rat . Read more (PubMed: 22135067) »

See 1 Publication for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Heart)
Specification Heart
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization No
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Username

Mr. Carl Hobbs

Verified customer

投稿 Feb 08 2013

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...

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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...

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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...

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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...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (hESC-derived cardiomyocyte)
Specification hESC-derived cardiomyocyte
Fixative Paraformaldehyde
Permeabilization 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
Username

Dr. David Anderson

Verified customer

投稿 Dec 03 2010

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"