Anti-Topoisomerase I/TOP1 Antibody

• Catalog Number: orb402201
• Category: Antibodies
• Description: Anti-Topoisomerase I/TOP1 Antibody. Tested in ELISA, IF, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, ICC, IF, WB
• Reactivity: Human, Mouse, Rat
• Isotype: Rabbit IgG
• Immunogen: E. coli-derived human Topoisomerase I recombinant protein (Position: E526-F765).
• 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: 100 kDa
• UniProt ID: P11387
• 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: DNA topoisomerase 1; DNA topoisomerase I; TOP1
• Note: For research use only
• Application notes: Western blot, 0.1-0.5μg/ml Immunocytochemistry/Immunofluorescence, 2μg/ml ELISA, 0.1-0.5μg/ml. Add 0.2ml of distilled water will yield a concentration of 500ug/ml
• Expiration Date: 12 months from date of receipt.

IF analysis of Topoisomerase I using anti-Topoisomerase I antibody. Topoisomerase I was detected in immunocytochemical section of Hela cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2 µg/mL rabbit anti-Topoisomerase I Antibody overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Western blot analysis of Topoisomerase I using anti-Topoisomerase I 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 HEL whole cell lysates, Lane 2: human Daudi whole cell lysates, Lane 3: human MOLT-4 whole cell lysates, Lane 4: human MCF-7 whole cell lysates, Lane 5: rat brain tissue lysates, Lane 6: mouse brain tissue 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-Topoisomerase I 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 Topoisomerase I at approximately 100 kDa. The expected band size for Topoisomerase I is at 91 kDa.