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Hcn2 Rabbit Polyclonal Antibody
Description
Research Area
Images & Validation
−| Tested Applications | ELISA, FC, IHC, WB |
|---|---|
| Dilution Range | Western blot, 0.25-0.5 μg/ml, Mouse, Rat Immunohistochemistry (Paraffin-embedded Section), 2-5 μg/ml, Mouse, Rat Flow Cytometry (Fixed), 1-3 μg/1x10^6 cells, Mouse ELISA, 0.1-0.5 μg/ml, - |
| Reactivity | Mouse, Rat |
Key Properties
−| Antibody Type | Primary Antibody |
|---|---|
| Host | Rabbit |
| Clonality | Polyclonal |
| Isotype | IgG |
| Immunogen | E.coli-derived mouse Hcn2 recombinant protein (Position: E660-L863). Mouse Hcn2 shares 87.4% and 97.5% amino acid (aa) sequence identity with human and rat HCN2, respectively. |
| Target | Potassium/sodium hyperpolarization-activated cyclic nucleotide-gated channel 2 |
| Molecular Weight | 110 kDa |
| Purification | Immunogen affinity purified. |
| Conjugation | Unconjugated |
Storage & Handling
−| Storage | Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles. |
|---|---|
| Form/Appearance | Lyophilized |
| Buffer/Preservatives | Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4. |
| Concentration | Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml. |
| Expiration Date | 12 months from date of receipt. |
| Disclaimer | For research use only |
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Flow Cytometry analysis of Neuro-2a cells using anti-Hcn2 antibody. Overlay histogram showing Neuro-2a cells (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-Hcn2 Antibody (1 µg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10 µg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 µg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Flow Cytometry analysis of NIH/3T3 cells using anti-Hcn2 antibody. Overlay histogram showing NIH/3T3 cells (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-Hcn2 Antibody (1 µg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10 µg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 µg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

IHC analysis of Hcn2 using anti-Hcn2 antibody. Hcn2 was detected in a paraffin-embedded section of mouse brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml rabbit anti-Hcn2 Antibody overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.

IHC analysis of Hcn2 using anti-Hcn2 antibody. Hcn2 was detected in a paraffin-embedded section of mouse cardiac tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml rabbit anti-Hcn2 Antibody overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.

IHC analysis of Hcn2 using anti-Hcn2 antibody. Hcn2 was detected in a paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml rabbit anti-Hcn2 Antibody overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.

IHC analysis of Hcn2 using anti-Hcn2 antibody. Hcn2 was detected in a paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml rabbit anti-Hcn2 Antibody overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.

IHC analysis of Hcn2 using anti-Hcn2 antibody. Hcn2 was detected in a paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml rabbit anti-Hcn2 Antibody overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.

IHC analysis of Hcn2 using anti-Hcn2 antibody. Hcn2 was detected in a paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml rabbit anti-Hcn2 Antibody overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.

IHC analysis of Hcn2 using anti-Hcn2 antibody. Hcn2 was detected in a paraffin-embedded section of rat cardiac tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml rabbit anti-Hcn2 Antibody overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.

Western blot analysis of Hcn2 using anti-Hcn2 antibody. Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions. Lane 1: rat brain tissue lysates, Lane 2: mouse brain tissue lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Hcn2 antigen affinity purified polyclonal antibody at 0.5 µg/mL overnight at 4°C, then washed with TBS-0.1% Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for Hcn2 at approximately 110 kDa. The expected band size for Hcn2 is at 97-110 kDa.
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Hcn2 Rabbit Polyclonal Antibody (orb1819380)
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