Anti-ALKBH5 Antibody

• Catalog Number: orb654309
• Category: Antibodies
• Description: Anti-ALKBH5 Antibody. Tested in ELISA, IF, ICC, WB applications. This antibody reacts with Human.
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, ICC, IF, WB
• Reactivity: Human
• Isotype: Rabbit IgG
• Immunogen: E.coli-derived human ALKBH5 recombinant protein (Position: E81-R180).
• 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: 52 kDa
• UniProt ID: Q6P6C2
• 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: RNA demethylase ALKBH5; Alkylated DNA repair prote
• Note: For research use only
• Application notes: Western blot, 0.1-0.25μg/ml, Human Immunocytochemistry/Immunofluorescence, 5μg/ml, Human 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 ALKBH5 using anti-ALKBH5 antibody and anti-Tubulin Alpha antibody. ALKBH5 was detected in immunocytochemical section of A549 cell. 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-ALKBH5 Antibody and mouse anti-Tubulin Alpha antibody overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG and DyLight®488 Conjugated Goat Anti-Mouse IgG were used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Western blot analysis of ALKBH5 using anti-ALKBH5 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 293T whole cell lysates, Lane 2: human Jurkat whole cell lysates, Lane 3: human Hela whole cell lysates, Lane 4: human HepG2 whole cell lysates, Lane 5: human U87 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-ALKBH5 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 ALKBH5 at approximately 52 kDa. The expected band size for ALKBH5 is at 44 kDa.