Anti-HNF-4-alpha Antibody (monoclonal, 6C8E9)

• Catalog Number: orb1145784
• Category: Antibodies
• Description: Anti-HNF-4-alpha Antibody (monoclonal, 6C8E9). Tested in IHC, WB applications. This antibody reacts with Human, Rat.
• Species/Host: Mouse
• Clonality: Monoclonal
• Clone Number: 6C8E9
• Tested applications: IHC, WB
• Reactivity: Human, Rat
• Isotype: Mouse IgG2a
• Immunogen: E.coli-derived human HNF-4-alpha recombinant protein (Position: Q164-I474). Human HNF-4-alpha shares 95% and 96% amino acid (aa) sequence identity with mouse and rat HNF-4-alpha, respectively.
• Antibody Type: Primary Antibody
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 53 kDa
• UniProt ID: P41235
• 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.
• Alternative names: AHUS2 antibody; Antigen defined by monoclonal anti
• Note: For research use only
• Application notes: Western blot, 0.25-0.5 μg/ml, Human, Rat Immunohistochemistry(Paraffin-embedded Section), 2-5 μg/ml, Rat. Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml
• Expiration Date: 12 months from date of receipt.

IHC analysis of HNF-4-alpha using anti-HNF-4-alpha antibody. HNF-4-alpha was detected in a paraffin-embedded section of rat colon 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 mouse anti-HNF-4-alpha Antibody overnight at 4°C. Peroxidase Conjugated Goat Anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Mouse IgG Super Vision Assay Kit with DAB as the chromogen.

IHC analysis of HNF-4-alpha using anti-HNF-4-alpha antibody. HNF-4-alpha was detected in a paraffin-embedded section of rat liver 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 mouse anti-HNF-4-alpha Antibody overnight at 4°C. Peroxidase Conjugated Goat Anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Mouse IgG Super Vision Assay Kit with DAB as the chromogen.

Western blot analysis of HNF-4-alpha using anti-HNF-4-alpha 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: human HepG2 whole cell lysates, Lane 2: rat liver 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 mouse anti-HNF-4-alpha antigen affinity purified monoclonal 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-mouse IgG-HRP secondary antibody at a dilution of 1:10000 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 HNF-4-alpha at approximately 53 kDa. The expected band size for HNF-4-alpha is at 53 kDa.