Anti-SIRT1 Antibody

• Catalog Number: orb215913
• Category: Antibodies
• Description: Anti-SIRT1 Antibody. Tested in IF, ICC, WB applications. This antibody reacts with Human.
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ICC, IF, WB
• Reactivity: Human
• Isotype: Rabbit IgG
• Immunogen: E.coli-derived human SIRT1 recombinant protein (Position: R112-K311). Human SIRT1 shares 90% amino acid (aa) sequence identity with mouse SIRT1.
• 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: 120 kDa
• UniProt ID: Q96EB6
• 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: NAD-dependent protein deacetylase sirtuin-1; hSIRT
• Note: For research use only
• Application notes: Western blot, 0.1-0.5μg/ml, Human Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human. Add 0.2ml of distilled water will yield a concentration of 500ug/ml
• Expiration Date: 12 months from date of receipt.

IF analysis of SIRT1 using anti-SIRT1 antibody. SIRT1 was detected in an immunocytochemical section of A431 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 5 µg/mL rabbit anti-SIRT1 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 SIRT1 using anti-SIRT1 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 HepG2 whole cell lysates, Lane 2: human MCF-7 whole cell lysates, Lane 3: huamn SiHa whole cell lysates, Lane 4: huamn Daudi whole cell lysates, Lane 5: huamn K562 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-SIRT1 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 SIRT1 at approximately 120 kDa. The expected band size for SIRT1 is at 82 kDa.