Anti-Fibroblast activation protein, alpha/FAP Antibody

• Catalog Number: orb1290013
• Category: Antibodies
• Description: Anti-Fibroblast activation protein, alpha/FAP Antibody. Tested in ELISA, Flow Cytometry, WB applications. This antibody reacts with Human.
• Clonality: Polyclonal
• Species/Host: Rabbit
• Isotype: Rabbit IgG
• Conjugation: Unconjugated
• Reactivity: Human
• Form/Appearance: Lyophilized
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Immunogen: E.coli-derived human Fibroblast activation protein, alpha/FAP recombinant protein (Position: Q65-Q580).
• UniProt ID: Q12884
• MW: 97 kDa
• Tested applications: ELISA, FC, WB
• Application notes: Western blot, 0.1-0.25 μg/ml, Human Flow Cytometry (Fixed), 1-3 μg/1x106 cells, Human ELISA, 0.1-0.5 μg/ml, -. Adding 0.2 ml of distilled water will yield a concentration of 500 μg/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: Prolyl endopeptidase FAP
• Note: For research use only

Flow Cytometry analysis of SiHa cells using anti-Fibroblast activation protein, alpha/FAP antibody. Overlay histogram showing SiHa cells (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Fibroblast activation protein, alpha/FAP Antibody (1 µg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10 µg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 µg/1x10^6) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Western blot analysis of Fibroblast activation protein, alpha/FAP using anti-Fibroblast activation protein, alpha/FAP 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 SiHa whole cell lysates, Lane 2: human U20S whole cell lysates, Lane 3: human Hela whole cell lysates, Lane 4: human U251 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-Fibroblast activation protein, alpha/FAP antigen affinity purified polyclonal antibody at 0.25 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 Fibroblast activation protein, alpha/FAP at approximately 97 kDa. The expected band size for Fibroblast activation protein, alpha/FAP is at 95 kDa.