Anti-Perforin/PRF1 Antibody

• Catalog Number: orb215909
• Category: Antibodies
• Description: Anti-Perforin/PRF1 Antibody. Tested in WB applications. This antibody reacts with Human.
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: WB
• Reactivity: Human
• Isotype: Rabbit IgG
• Immunogen: E.coli-derived human Perforin recombinant protein (Position: E175-W555). Human Perforin shares 68% amino acid (aa) sequence identity with both mouse and rat Perforin.
• Antibody Type: Primary Antibody
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 61 kDa
• UniProt ID: P14222
• 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: Perforin-1; P1; Cytolysin; Lymphocyte pore-forming
• Note: For research use only
• Application notes: Western blot, 0.1-0.5μg/ml, Human. 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 Perforin using anti-Perforin antibody. Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. Lane 1: recombinant human Perforin protein 0.5 ng. 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-Perforin 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 Perforin at approximately 40 kDa. The expected band size for Perforin is at 40 kDa.

Western blot analysis of Perforin using anti-Perforin 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 Hela whole cell lysates, Lane 2: human COLO320 whole cell lysates, Lane 3: human HepG2 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-Perforin 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 Perforin at approximately 61 kDa. The expected band size for Perforin is at 61 kDa.