|Application ||WB, IP|
|Reactivity||Human, Mouse, Rat|
|Calculated MW||129042 Da|
|Homology||Rabbit - identical; rat - 35/37 amino acid residues identical.|
|Other Names||Potassium/sodium hyperpolarization-activated cyclic nucleotide-gated channel 4, HCN4|
|Related products for control experiments||Control peptide antigen (supplied with the antibody free of charge).|
|Target/Specificity||GST fusion protein with sequence HGHLHDSAEERRLIAEGDASPG EDRTPPGLAAEPERP, corresponding to amino acid residues 119-155 of human HCN4 (Accession Q9Y3Q4) , (MW: 31 kDa.).ֲ ֲ Intracellular, N-terminus.|
|Peptide Confirmation||Confirmed by DNA sequence and SDS PAGE.|
|Application Details||Western blot analysis (WB): - Rat heart lysate (1:200) (see see Atkinson, A.J. et al. (2013) in Product Citations). - Human HCN4 transfected in HEK-293T cells. (see Nof, E. et al. (2007) in Product Citations.). Immunoprecipitation (IP): - Rat thalamus lysates (see Whitaker, G.M. et al. (2007) in Product Citations.). Immunocytochemistry (IC): - HEK-293 transfected cells (1:200) (see Altomare, C. et al. (2003) in Product Citations.). Immunohistochemistry (IH): - Mouse heart sections (see Mahida, S. et al. (2014) in Product Citations). - Rat heart sections (1:20) see (see Atkinson, A.J. et al. (2013) in Product Citations). - Mouse eye sections (1:500) (see Zhang, J. et al. (2013) in Product Citations.).|
|Format||Affinity purified antibody, lyophilized powder|
|Reconstitution||25 µl, 50 µl or 0.2 ml deionized water, depending on the sample size.|
|Antibody Concentration After Reconstitution||0.8 mg/ml.|
|Storage Before Reconstitution||Lyophilized powder can be stored intact at room temperature for several weeks. For longer periods, it should be stored at -20°C.|
|Storage After Reconstitution||The reconstituted solution can be stored at 4ºC for up to 2 weeks. For longer periods, small aliquots should be stored at -20ºC or below. Avoid multiple freezing and thawing. The further dilutions should be made using a carrier protein such as BSA (1%). Centrifuge all antibody preparations before use (10000 × g 5 min).|
|Control Antigen Storage Before Reconstitution||Lyophilized powder can be stored intact at room temperature for several weeks. For longer periods, it should be stored at -20°C.|
|Control Antigen Reconstitution||100 µl PBS.|
|Control Antigen Storage After Reconstitution||-20ºC.|
|Preadsorption Control||3 µg fusion protein per 1 µg antibody.|
|Formulation||Lyophilized powder. Phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN3.|
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Provided below are standard protocols that you may find useful for product applications.
Hyperpolarization-activated cation currents (Ih) appear in the heart and the brain and have a 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 is comprised of four homologous members, HCN1-4. Each HCN subunit consists of six transmembrane domains (TM), a pore region between TM5-TM6 and a binding domain for 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 a homology of about 60%. However, their similarity decreases in the cytoplasmic N- and C-termini. The HCN1-4 channels mainly differ from each other in their speed of activation and the extent to which they are modulated by cAMP. HCN1, weakly affected by cAMP, is the fastest channel, followed by HCN2, HCN3 and HCN4. HCN4 is highly expressed in a restricted manner in adult sinoatrial (SA) node, constituting a good molecular marker for the adult cardiac pacemaker and might serve as a unique marker of the developing SA node .4,5 mRNA expression of HCN4 is most abundant in medial habenula and anterior and principal relay nuclei of the thalamus.6
References 1. Much, B. et al (2003) J.Biol.Chem. 278, 43781. 2. Notomi, T. and Shigemoto R. (2004) J. Comp. Neurol. 471, 241. 3. Gravante, B. et al (2004) J.Biol.Chem. 279, 43847. 4. Moosmang, S. et al. (2001) Eur. J. Biochem. 268, 1646. 5. Garcia-Frigola, C. et al. (2003) Gene Expr Patterns 3, 777. 6. Monteggia, L. et al. (2000) Molec. Brain Res. 1-2, 129.
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