All rats were deeply anesthetized with sodium pentobarbital (60 mg/kg), and the L4-L6 spinal cord was quickly harvested and stored at −80°C before being subjected to the following procedure. Samples were homogenized in Tris buffer with a cocktail of proteinase inhibitors and phosphatase inhibitors and were subjected to SDS-PAGE, followed by electrophoretic transfer onto polyvinylidene fluoride (PVDF) membranes. The membranes were placed in the block buffer at room temperature for 1 h and incubated with one of the following primary antibodies: anti-HCN2 (Proteintech, Wuhan, China), anti-NR2B (Proteintech), anti-pCaMKII (Cell Signalling Technology, USA); anti-CaMKII (Cell Signalling Technology); anti-pCREB (Cell Signalling Technology); anti-CREB (Cell Signalling Technology) at 4°C for 12 h, and then incubated with horseradish peroxidase-conjugated anti-rabbit IgG (Proteintech) for 1 h. Quantification of the band for each protein was performed with Image J software.
Anti hcn2
Manufactured by Proteintech
Sourced in United States
Anti-HCN2 is a primary antibody product that recognizes the HCN2 (hyperpolarization-activated cyclic nucleotide-gated potassium channel 2) protein. HCN2 is a subunit of the HCN channel, which plays a role in regulating neuronal excitability and the pacemaker activity of the heart.
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Lab products found in correlation
Anti nr2b, by Proteintech (1 mentions)
Anti p camkii, by Cell Signaling Technology (1 mentions)
Anti camkii, by Cell Signaling Technology (1 mentions)
Horseradish peroxidase conjugated anti rabbit igg, by Proteintech (1 mentions)
Anti pcreb, by Cell Signaling Technology (1 mentions)
Anti creb, by Cell Signaling Technology (1 mentions)
Goat anti rabbit igg secondary antibody, by Thermo Fisher Scientific (1 mentions)
Ibright 1500 imaging system, by Thermo Fisher Scientific (1 mentions)
Anti β actin, by Proteintech (1 mentions)
2 protocols using anti hcn2
1
Spinal Cord Protein Expression Analysis
2
Western Blot Analysis of HCN1 and HCN2
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After the behavioral test, fresh VPL tissue was homogenized in ice-cold radio immunoprecipitation assay lysis buffer and 1% protease inhibitor for 1 h and then centrifuged at 12,000 r/min for 15 min at 4 °C. For electrophoretic blotting, samples were separated by 8% sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to a nitrocellulose membrane. Subsequently, the membrane was blocked at room temperature for 1 h and incubated with rabbit polyclonal anti-HCN1 (1:1000; Proteintech, IL, USA) and anti-HCN2 (1:1000; Proteintech) or rabbit polyclonal anti-β-actin (1:1000; Proteintech) antibodies at 4 °C overnight. After 3 washes, the membranes were incubated with goat anti-rabbit IgG secondary antibody (1:2000; Thermo Fisher Scientific, MA, USA) for 2 h at room temperature. The immunoreactive bands were visualized by an iBright 1500 imaging system (Thermo Fisher Scientific) and quantified by using ImageJ software (http://rsb.info.nih.gov/ij/ ).
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