Overview
- Peptide (C)EEAGPAGEPRGSQAS, corresponding to amino acid residues 147-161 of human HCN2 (Accession Q9UL51). Intracellular, N-terminus.
Western blot analysis of rat brain membranes:1. Anti-HCN2 Antibody (#APC-030), (1:200).
2. Anti-HCN2 Antibody, preincubated with HCN2 Blocking Peptide (#BLP-PC030).
- Rat thalamus lysate (Whitacker, G.M. et al. (2007) J. Biol. Chem. 282, 22900.).
Expression of HCN2 in rat cerebellumImmunochistochemical staining of rat cerebellum frozen sections using Anti-HCN2 Antibody (#APC-030 ). A. HCN2 (red) appears in Purkinje cells (arrows). B. Staining of astrocytes with mouse anti-glial fibrillary acidic protein (GFAP, green demonstrates the restriction of HCN2 to neuronal cell bodies. C. Confocal merge of HCN2 and GFAP images demonstrates the respective localization of these proteins.
Expression of HCN2 in mouse hypothalamusImmunohistochemical staining of mouse hypothalamus using Anti-HCN2 Antibody (#APC-030). A. HCN2 (red) appears in cells of the paraventricular nucleus (PVN, arrows). B. Staining of paraventricular nerve cells with mouse anti-calcium binding protein (CBD28k, green). C. Confocal merge of HCN2 and CBD28k demonstrates some co-localization. V = Third ventricle.
Colocalization of HCN4 and HCN2 in mouse thalamusImmunohistochemical staining of mouse thalamus frozen section using Guinea pig Anti-HCN4 Antibody (#APC-052-GP) 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.
Expression of HCN2 in rat DRG primary cultureImmunocytochemical staining of paraformaldehyde-fixed and permeabilized rat dorsal root ganglion (DRG) primary culture using Anti-HCN2 Antibody (#APC-030), (1:100), (green). Cells were stained with Anti-HCN2 Antibody followed by goat anti-rabbit-AlexaFluor-488 secondary antibody. Nuclear staining of cells using the cell-permeable DNA dye Hoechst 33342 (blue).
- Much, B. et al. (2003) J. Biol. Chem. 278, 43781.
- Notomi, T. et al. (2004) J. Comp. Neurol. 471, 241.
- Hofmann, F. et al. (2005) Pharmacol. Rev. 57, 455.
Hyperpolarization-activated cation currents (Ih) appear in the heart and the brain having 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 comprises four homologous members, named HCN1-4. Each HCN subunit consist of six transmembrane domains (TM), a pore region between TM5-TM6 and a binding domain to 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 homology of about 60%. However, they are diverging from each other in their cytoplasmic N- and the C-terminus. The channels HCN1-4 mainly differ from each other with regard to their speed of activation and the extent by which they are modulated by cAMP. HCN1 is the fastest channel, followed by HCN2, HCN3 and HCN4.2,3
HCN2 is the most abundant neuronal channel and is found almost ubiquitously in the brain.3
Anti-HCN2 Antibody (#APC-030) is a highly specific antibody directed against an epitope of the human protein. The antibody can be used in western blot, immunoprecipitation, immunohistochemistry, and immunocytochemistry applications. It has been designed to recognize HCN2 from human, rat, and mouse samples.
Knockout validation of Anti-HCN2 Antibody in mouse brain.Immunohistochemical staining of mouse coronal brain sections using Anti-HCN2 Antibody (#APC-030). HCN2 staining (red) is broadly detected across the coronal section. Lack of HCN2 staining is observed in HCN2-/- mice. Adapted from Hammelmann, V. et al. (2011) PLoS ONE 6, e17078.
Applications
Citations
Powered by Bioz- Immunohistochemical staining of mouse brain sections. Tested in HCN2-/- mice.
Hammelmann, V. et al. (2011) PLoS ONE 6, e17078. - Immunohistochemical staining of mouse heart sections. Tested in HCN2-/- mice.
Herrmann, S. et al. (2011) J. Mol. Cell. Cardiol. 51, 997.
- Human sinoatrial node lysate (1:500).
Li, N. et al. (2015) Circ. Arrhythm. Electrophysiol. 8, 1219.
- Rat thalamus lysate.
Whitacker, G.M. et al. (2007) J. Biol. Chem. 282, 22900.
- Rat diaphragm sections (1:25).
Negrini, D. et al. (2016) Am. J. Physiol. 311, H892. - Human sinoatrial node sections (1:100).
Li, N. et al. (2015) Circ. Arrhythm. Electrophysiol. 8, 1219. - Mouse vestibular ganglia.
Horwitz, G.C. et al. (2014) J. Gen. Physiol. 143, 481. - Mouse brain sections. Also tested in HCN2-/- mice.
Hammelmann, V. et al. (2011) PLoS ONE 6, e17078. - Mouse heart sections. Also tested in HCN2-/- mice.
Herrmann, S. et al. (2011) J. Mol. Cell. Cardiol. 51, 997.
- Cao Ehlker, X. et al. (2013) J. Biol. Chem. 288, 7580.
- Nakashima, N. et al. (2013) J. Physiol. 591, 1749.
- Rusznak, Z. et al. (2013) Eur. J. Neurosci. 37, 876.
- Weisbrod, D. et al. (2013) Proc. Natl. Acad. Sci. U.S.A. 110, E1685.
- Ramakrishnan, N.A. et al. (2012) J. Biol. Chem. 287, 37628.
- Doan, T.N. et al. (2004) J. Neurosci. 24, 3335.
- Jiang, M. et al. (2004) Circulation 109, 1783.
- Macri, V. and Accili, A.A. (2004) J. Biol. Chem. 279, 16832.
- Much, B. et al. (2003) J. Biol. Chem. 278, 43781.
- Han, W. et al. (2002) Circ. Res. 91, 790.
- Moroni A. et al. (2001) J. Biol. Chem. 276, 29233.
