CD13/ANPEP Antibody (monoclonal, 9G5)

• Catalog Number: orb763149
• Category: Antibodies
• Description: CD13/ANPEP Antibody (monoclonal, 9G5)
• Species/Host: Mouse
• Clonality: Monoclonal
• Clone Number: 9G5
• Tested applications: ICC, IF, IHC, WB
• Reactivity: Human, Monkey, Mouse, Rat
• Isotype: Mouse IgG2b
• Immunogen: E.coli-derived human CD13/ANPEP recombinant protein (Position: D148-S966).
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 150 kDa
• UniProt ID: P15144
• 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.
• Note: For research use only
• Application notes: Tested Species: In-house tested species with positive results. Other applications have not been tested. Optimal dilutions should be determined by end users. Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Expiration Date: 12 months from date of receipt.

Western blot analysis of CD13/ANPEP using anti-CD13/ANPEP antibody (orb763149). 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 THP-1 whole cell lysates, Lane 2: human PC-3 whole cell lysates, Lane 3: monkey kidney tissue lysates, Lane 4: rat kidney tissue lysates, Lane 5: rat liver tissue lysates, Lane 6: mouse kidney 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 mouse anti-CD13/ANPEP antigen affinity purified monoclonal antibody (Catalog # orb763149) 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-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # orb90502) with Tanon 5200 system. A specific band was detected for CD13/ANPEP at approximately 150 kDa. The expected band size for CD13/ANPEP is at 150 kDa.

IHC analysis of CD13/ANPEP using anti-CD13/ANPEP antibody (orb763149). CD13/ANPEP was detected in a paraffin-embedded section of rat intestine tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml mouse anti-CD13/ANPEP Antibody (orb763149) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90443) with DAB as the chromogen.

IF analysis of CD13/ANPEP using anti-CD13/ANPEP antibody (orb763149). CD13/ANPEP was detected in an immunocytochemical section of HepG2 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (orb90553) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL mouse anti-CD13/ANPEP Antibody (orb763149) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse 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.