Label ATTO-594. Maximum absorption 601 nm; maximum fluorescence 627 nm. The fluorescence is excited most efficiently in the 580 - 615 nm range. This label is related to the Rhodamine dyes and can be used with filters used to detect Texas Red and Alexa-594.
Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
Reconstitution 50 µl double distilled water (DDW).
Antibody concentration after reconstitution 1 mg/ml.
Storage after reconstitution The reconstituted solution can be stored at 4°C, protected from the light, 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 × g 5 min).
Standard quality control of each lot Western blot analysis (unlabeled antibody, #APC-030), and immunohistochemistry (labeled antibody).
Applications: if, ih
May also work in: ic*
Immunohistochemistry
Rat cerebellum.
Expression of HCN2 in rat cerebellum
Immunohistochemical staining of rat cerebellum using Anti-HCN2-ATTO Fluor-594Antibody (#APC-030-AR). A. HCN2 staining (red) appears in Purkinje neurons. B. Nuclear staining using DAPI as the counterstain (Blue). C. Merged images of A and B.
Scientific background
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
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
Expression of HCN2 in rat cerebellumImmunohistochemical staining of rat cerebellum using Anti-HCN2-ATTO Fluor-594 Antibody (#APC-030-AR). A. HCN2 staining (red) appears in Purkinje neurons. B. Nuclear staining using DAPI as the counterstain (Blue). C. Merged images of A and B.
