HIF-1 alpha/HIF1A Antibody

• Catalog Number: orb570389
• Category: Antibodies
• Description: HIF-1 alpha/HIF1A Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, WB
• Reactivity: Human
• Isotype: Rabbit IgG
• Immunogen: E.coli-derived human HIF-1 alpha/HIF1A recombinant protein (Position: H197-R718).
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Dilution range: Western blot, 0.25-0.5μg/ml, Human Direct ELISA, 0.1-0.5μg/ml, Human
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 123 kDa
• UniProt ID: Q16665
• 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: Hypoxia-inducible factor 1-alpha; HIF-1-alpha; HIF
• Note: For research use only
• Application notes: Tested Species: In-house tested species with positive results. 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 HIF1A using anti-HIF1A antibody (orb570389). 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 U-87MG whole cell lysates, Lane 2: human U2OS whole cell lysates, Lane 3: human PC-3 whole cell lysates, Lane 4: human A549 whole cell lysates, Lane 5: human HepG2 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-HIF1A antigen affinity purified polyclonal antibody (Catalog # orb570389) 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 HIF1A at approximately 123KD. The expected band size for HIF1A is at 93KD.