Overview

  • Product nameAnti-HCN4 antibody [SHG 1E5]
    See all HCN4 primary antibodies
  • Description
    Rat monoclonal [SHG 1E5] to HCN4
  • Tested applicationsSuitable for: WB, ICC/IF, IHC-Fr, Flow Cyt, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Chimpanzee
  • Immunogen

    Synthetic peptide: SHGSLLLPPA SSPPPPQVPQ RRGTPPLTPG RLTQDLKL, corresponding to amino acids 1048-1085 of Human HCN4.

Properties

Applications

Our Abpromise guarantee covers the use of ab32675 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 1/5000. Detects a band of approximately 132 kDa (predicted molecular weight: 130 kDa).
ICC/IF Use at an assay dependent concentration. PubMed: 19775764
IHC-Fr 1/1000.
Flow Cyt 1/50.

ab18407 - Rat monoclonal IgG1, is suitable for use as an isotype control with this antibody.

IHC-P 1/10 - 1/100.

Target

  • RelevanceHCN4 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 localizationMembrane; 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 [SHG 1E5] images

  • Anti-HCN4 antibody [SHG 1E5] (ab32675) at 1/200 dilution + Rat Eye Lysate at 25 µg

    Secondary
    HRP Conjugate

    Predicted band size : 130 kDa
    Observed band size : 132 kDa (why is the actual band size different from the predicted?)
  • Overlay histogram showing PC12 cells stained with ab32675 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32675, 1/50 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rat IgG (H+L) (ab98386) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rat IgG1 [RTK2071] (ab18412, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in PC12 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween used under the same conditions.

  • Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human tonsil tissue tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:20 with a rat monoclonal antibody recognizing HCN4 ab32675 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

  • Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human heart tissue tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:20 with a rat monoclonal antibody recognizing HCN4 ab32675 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

References for Anti-HCN4 antibody [SHG 1E5] (ab32675)

This product has been referenced in:
  • Sun C  et al. The short stature homeobox 2 (Shox2)-bone morphogenetic protein (BMP) pathway regulates dorsal mesenchymal protrusion development and its temporary function as a pacemaker during cardiogenesis. J Biol Chem 290:2007-23 (2015). Mouse . Read more (PubMed: 25488669) »
  • Lescroart F  et al. Early lineage restriction in temporally distinct populations of Mesp1 progenitors during mammalian heart development. Nat Cell Biol 16:829-40 (2014). Read more (PubMed: 25150979) »

See all 9 Publications for this product

Product Wall

Thank you for your response.

"No primary control" means that you perform the staining protocol as normal (fixation, blocking, secondary antibody, detection) - except you need to omit the incubation step with the primary antibody. Ins...

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Thank you for your enquiry and your interest in our products.

The immunogen used to raise this antibody was a synthetic peptide: SHGSLLLPPA SSPPPPQVPQ RRGTPPLTPG RLTQDLKL, corresponding to amino acids 1048-1085 of Human HCN4:SHGSLLLPPASSPPPPQV...

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DISCOUNT CODES:
Ab109235: ***
Ab32675: ***

Expiration date: October 5, 2012

Thesecodes will each give you 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview for each antibody...

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Vielen Dank dafür, dass meine Fragen zu beantworten.


Ich möchte darauf kurz hinweisen, dass alle Antikörper eine abgestimmte Optimierung benötigen. Es ist daher nichts Ungewöhnliches ein paar Optimierungsexperimente mit einem neuen Ant...

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Es tut mir leid, dass Sie Probleme mit unserem Antikörper hatten.

Ich habe da noch ein zwei Fragen bezüglich des Protokolls:

1. Wie lange haben Sie die Antigen Maskierung durchgeführt? Haben Sie eine Zeitreihe durchgeführt?
...

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Thank you very much for your interest in ab32675.

To our knowledge,ab32675 has not been tested in immunoprecipitation (IP). Therefore, I can offer a discount off a future purchase if you buy ab32675 now, test it inIP and submit feedback to u...

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Thank you for your enquiry. Further to your telephone enquiry I have been in correspondence with the source of this antibody. Unfortunately the immunogen peptide has been exhausted for this antibody and therefore unavailable for purchase. The synth...

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