HIF-2-alpha/EPAS1 Antibody

• Catalog Number: orb96817
• Category: Antibodies
• Description: HIF-2-alpha/EPAS1 Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: WB
• Predicted Reactivity: Hamster
• Reactivity: Human, Mouse, Rat
• Isotype: Rabbit IgG
• Immunogen: A synthetic peptide corresponding to a sequence in the middle region of human HIF-2-alpha (282-301aa, HALDSENMTKSHQNLCTKGQ), identical to the related mouse and rat sequence.
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Dilution range: Western blot, 0.1-0.5μg/ml, Human, Mouse, Rat
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 96718 MW
• UniProt ID: Q9JHS1
• Storage: Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
• Alternative names: Endothelial PAS domain-containing protein 1;EPAS-1
• Note: For research use only
• Application notes: Tested Species: In-house tested species with positive results. Predicted Species: Species predicted to be fit for the product based on sequence similarities. Other applications have not been tested. Optimal dilutions should be determined by end users. . Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
• Expiration Date: 12 months from date of receipt.

Western blot analysis of HIF2A using anti-HIF2A antibody (orb96817). 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 50ug of sample under reducing conditions. Lane 1: rat thymus tissue lysates, Lane 2: rat testis tissue lysates, After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-HIF2A antigen affinity purified polyclonal antibody (Catalog # orb96817) 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 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # orb90503) with Tanon 5200 system. A specific band was detected for HIF2A at approximately 110-120KD. The expected band size for HIF2A is at 96KD.

Western blot analysis of HIF-2A using anti-HIF-2A antibody (orb96817). 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 50ug of sample under reducing conditions. Lane 1: human MCF-7 whole cell lysates, Lane 2: human HeLa whole cell lysates, Lane 3: human Jurkat whole cell lysates, After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-HIF-2A antigen affinity purified polyclonal antibody (Catalog # orb96817) 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 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # orb90503) with Tanon 5200 system. A specific band was detected for HIF-2A at approximately 120KD. The expected band size for HIF-2A is at 96KD.

Western blot analysis of HIF-2A using anti-HIF-2A antibody (orb96817). 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 50ug of sample under reducing conditions. Lane 1: mouse thymus tissue lysates, Lane 2: mouse lun tissue lysates, After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-HIF-2A antigen affinity purified polyclonal antibody (Catalog # orb96817) 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 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # orb90503) with Tanon 5200 system. A specific band was detected for HIF-2A at approximately 120KD. The expected band size for HIF-2A is at 96KD.

Western blot analysis of HIF-2A using anti-HIF-2A antibody (orb96817). 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 50ug of sample under reducing conditions. Lane 1: rat brain tissue lysates, Lane 2: rat RH35 whole cell lysates, Lane 3: rat small intestine tissue lysates, After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-HIF-2A antigen affinity purified polyclonal antibody (Catalog # orb96817) 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 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # orb90503) with Tanon 5200 system. A specific band was detected for HIF-2A at approximately 120KD. The expected band size for HIF-2A is at 96KD.