Heme Oxygenase 1/HMOX1 Rabbit Polyclonal Antibody

IHC analysis of HMOX1 using anti-HMOX1 antibody. HMOX1 was detected in a paraffin-embedded section of human tonsil 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-HMOX1 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 heme oxygenase 1/HMOX1 using anti-heme oxygenase 1/HMOX1 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. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 1x PBS +0.2% Tween-20 + 10% nonfat dry milk for 1.5 hour at RT. The membrane was incubated with rabbit anti-heme oxygenase 1/HMOX1 antibody at 0.5 µg/mL overnight at room temperature for 3 hours, then washed with TBS-0.1% Tween 3 times with 5 minutes each and probed with a HRP-conjugated anti rabbit antibodies at 4℃ for 12 hours. The signal is developed using an Enhanced chemiluminescence for one minute. A specific band was detected for heme oxygenase 1/HMOX1 at approximately 32 kDa. The expected band size for heme oxygenase 1/HMOX1 is at 32 kDa.

Western blot analysis of HMOX1 using anti-HMOX1 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 A549 whole cell lysates, Lane 2: human Hela whole cell 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-HMOX1 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 HMOX1 at approximately 33 kDa. The expected band size for HMOX1 is at 33 kDa.