Name: Anti-KCNH2 (HERG) (extracellular) Antibody
Brand: Alomone Labs
SKU: APC-109

Overview

Cat #: APC-109

Alternative Name K_V11.1, Potassium voltage-gated channel subfamily H member 2, Ether-a-go-go-related channel 1

Lyophilized Powder yes

Type: Polyclonal

Host: Rabbit

Reactivity: h, r

May also work in: m*

Immunogen

Peptide AFLLKETEEGPPATEC corresponding to residues 430-445 of human K_V11.1 (HERG) (Accession Q12809). Extracellular, between S1 and S2 domains.

Accession (Uniprot) Number Q12809

Gene ID 3757

Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.

Homology Rat, mouse - 11/16 amino acid residues identical; dog, rabbit - 15/16 amino residues identical.

RRID AB_2039913.

Purity Affinity purified on immobilized antigen.

Form Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN₃.

Form Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4.

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.

Reconstitution 0.2 ml double distilled water (DDW).

Antibody concentration after reconstitution 0.6 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).

Standard quality control of each lot Western blot analysis.

Applications: ic, if, ifc, ih, ip, lci, wb

Western blot

Western blot analysis of K_V11.1 (HERG)-transfected HEK-293 cells:
1. Anti-KCNH2 (HERG) (extracellular) Antibody (#APC-109), (1:200).
2. Anti-KCNH2 (HERG) (extracellular) Antibody, preincubated with KCNH2/HERG (extracellular) Blocking Peptide (#BLP-PC109).

Immunoprecipitation

Immunoprecipitation of K_V11.1-expressing HEK-293 cells:
1. Cell lysate.
2. Cell lysate + protein A beads + Anti-KCNH2 (HERG) (extracellular) Antibody (#APC-109).
3. Cell lysate + protein A beads + Anti-KCNH2 (erg1) Antibody (#APC-016).
4. Cell lysate + protein A beads + Anti-KCNH2 (HERG) Antibody (#APC-062).
5. Cell lysate + protein A beads + pre-immune rabbit serum.

Red arrow indicates K_V11.1 while the black arrow shows the IgG heavy chain.
Immunoblot was performed with Anti-KCNH2 (HERG) (extracellular) Antibody.

Immunohistochemistry

Expression of K_V11.1 (HERG) in rat heart
Immunohistochemical staining of rat heart using Anti-KCNH2 (HERG) (extracellular) Antibody (#APC-109). A. Transversal section of the atrium wall; note that arterial smooth muscle fibers were not stained (green arrow). B. Longitudinal section of the myocardium. C. Section showing myocardium and endocardium (red arrow). DAB product is brown and the counterstain is cresyl violet.

Indirect flow cytometry

Cell surface detection of K_V11.1 (HERG) by indirect flow cytometry in live intact human chronic myelogenous leukemia (K562) cells:
^___ Cells.
^___ Cells + goat-anti-rabbit-FITC.
^___ Cells + Anti-KCNH2 (HERG) (extracellular) Antibody (#APC-109), (2.5μg) + goat-anti-rabbit-FITC.
The blocking peptide is not suitable for this application.

Live cell imaging / Immunocytochemistry

Expression of K_V11.1 (HERG) in HEK-293 transfected cells
Cell surface detection of K_V11.1 in live intact HEK-293 transfected cells. Cells were stained with Anti-KCNH2 (HERG) (extracellular) Antibody (#APC-109), (1:25) (left panel) or with Anti-KCNH2 (HERG) (extracellular) Antibody preincubated with the blocking peptide (right panel), followed by goat anti-rabbit-AlexaFluor-555 secondary antibody (red). Nuclei of the living cells were stained with the cell permeable dye Hoechst 33342 (blue). Note that not all cells are stained, indicating that not all the K_V11.1 channels are expressed at the cell membrane at the moment of staining.

References

Curran, M.E. et al. (1995) Cell 80, 795.
Sanguinetti, M.C. et al. (1995) Cell 81, 299.
Pardo, L.A. et al. (2004) Physiology 19, 285.

Scientific background

The K_V11.1 (HERG) channel is a member of the ether-a-go-go (EAG) subfamily of voltage-dependent K⁺ channels that includes the related proteins K_V11.2 and K_V11.3 (erg2 and erg3). K_V11.1 possesses the signature structure of the voltage-dependent K⁺ channels: six membrane-spanning domains and intracellular N- and C-termini.

The K_V11.1 current is characterized by strong inward rectification with slow activation and very rapid inactivation kinetics. The channel is expressed in the brain and heart (where it underlies the I_Kr current) and has a central role in mediating repolarization of action potentials.^1,2

Mutations in the K_V11.1 channel cause inherited long QT syndrome (LQTS) or abnormalities in the repolarization of the heart that are associated with life-threatening arrhythmias and sudden death. All the identified K_V11.1 mutations produce loss of function of the channel via several cellular mechanisms ranging from alterations of gating properties, alterations of channel permeability/selectivity and alterations in intracellular channel trafficking that decreases the number of channels that reach the cell membrane.^1,2

Recently, drug-induced forms of LQTS have been reported for a wide range of non-cardiac drugs including antihistamines, psychoactive agents and antimicrobials. All these drugs potently block the K_V11.1 channel as an unintended side effect, prompting regulatory drug agencies to issue recommendations for the testing of new drugs for their potential K_V11.1 blocking effect.

In addition, K_V11.1 expression was found to be upregulated in several tumor cell lines of different histogenesis, suggesting that it confers the cells some advantage in cell proliferation. Indeed, in several studies it has been shown that inhibition of the K_V11.1 current leads to a decrease in tumor cell proliferation.³

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

Lyophilized Powder

Anti-KCNH2 (HERG) (extracellular) Antibody (#APC-109) is a highly specific antibody directed against an extracellular epitope of the human K_V11.1 channel. The antibody can be used in western blot, immunoprecipitation, immunohistochemical, immunocytochemical and indirect flow cytometry applications. It has been designed to recognize K_V11.1 from human, rat, and mouse samples.

Certificate of Analysis SDS

For research purposes only, not for human use

Last Update: 29/10/2024

Applications

Citations

More product citations

Crottes, D. et al. (2011) J. Biol. Chem. 286, 27947.
Mihic, A. et al. (2011) PLoS One. 6, e18273.
Smith, J.L. et al. (2011) Am. J. Physiol. Cell Physiol. 301, C75.
Stump, M.P. et al. (2011) Am. J. Physiol. Heart Circ. Physiol. 300, H312.
Wu, Z-Y. et al. (2011) J. Biol. Chem. 286, 22186.
Nof, E. et al. (2010) Circ. Cardiovasc. Genet 3, 199.
Delisle, B.P. et al. (2009) J. Biol. Chem. 284, 2844.
Potet, F. et al. (2009) J. Mol. Cell. Cardiol. 46, 257.
Chtcheglova, L.A. et al. (2008) Pflugers Arch. 456, 247.
Claassen, S. et al. (2008) J. Membr. Biol. 222, 31.
Takemasa, H. et al. (2008) Br. J. Pharmacol. 153, 439.
Wu, Z-Y. et al. (2008) Endocrinology 149, 5061.

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Anti-KCNH2 (HERG) (extracellular) Antibody

Overview

Application key:

Species reactivity key:

Applications

Citations

Specifications

Scientific Background

Resources

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