Anti-Osteopontin/SPP1 Antibody

[list_product_images]Figure 1. Western blot analysis of Osteopontin using anti- Osteopontin antibody (PB0589).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: mouse brain tissue lysates,Lane 3: human SHG-44 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- Osteopontin antigen affinity purified polyclonal antibody (Catalog # PB0589) 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 # EK1002) with Tanon 5200 system. A specific band was detected for Osteopontin at approximately 66,40KD. The expected band size for Osteopontin is at 35KD.|Figure 2. Western blot analysis of OPN using anti- OPN antibody (PB0589). 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 eachlane was loaded with 50ug of sample under reducing conditions.After Electrophoresis, proteins were transferred to a Nitrocellulosemembrane at 150mA for 60 minutes.Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-OPN antibody (Catalog # PB0589) at 0.5ug/ml 4℃ overnight, 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 (Catalog # EK1002) with Tanon 5200 system. Specific bands were detected for OPN at approximately 66KD (OPN precursor) and 40KD (OPN cleaved).[/list_product_images]