Anti-Osteopontin/SPP1 Antibody

• Catalog Number: orb308760
• Category: Antibodies
• Description: Anti-Osteopontin/SPP1 Antibody. Tested in ELISA, WB applications. This antibody reacts with Human, Mouse, Rat.
• Clonality: Polyclonal
• Species/Host: Rabbit
• Isotype: Rabbit IgG
• Conjugation: Unconjugated
• Reactivity: Human, Mouse, Rat
• Form/Appearance: Lyophilized
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Immunogen: A synthetic peptide corresponding to a sequence at the C-terminus of human Osteopontin, different from the related mouse sequence by eight amino acids, and from the related rat sequence by seven amino acids.
• UniProt ID: P10451
• MW: 66 kDa, 25-55 kDa
• Tested applications: ELISA, WB
• Application notes: Western blot, 0.1-0.5μg/ml, Human, Mouse, Rat, -ELISA, 0.1-0.5μg/ml, -. Add 0.2ml of distilled water will yield a concentration of 500ug/ml
• Antibody Type: Primary Antibody
• 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: Osteopontin; Bone sialoprotein 1; Nephropontin; Se
• Note: For research use only

Western blot analysis of Osteopontin using anti-Osteopontin 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 50 ug 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 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 with Tanon 5200 system. A specific band was detected for Osteopontin at approximately 66, 25-55 KD. The expected band size for Osteopontin is at 35 KD.