Anti-CD8 alpha/CD8A Antibody

• Catalog Number: orb234295
• Category: Antibodies
• Description: Anti-CD8 alpha/CD8A Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: IHC, WB
• Reactivity: Human
• Isotype: Rabbit IgG
• Immunogen: E.coli-derived human CD8 alpha recombinant protein (Position: S22-A187). Human CD8 alpha shares 49% and 47% amino acid (aa) sequence identity with mouse and rat CD8 alpha, respectively.
• 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: 55 kDa
• UniProt ID: P01732
• 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: T-cell surface glycoprotein CD8 alpha chain; T-lym
• Note: For research use only
• Application notes: Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, HumanWestern blot, 0.1-0.5μg/ml, Human. Add 0.2ml of distilled water will yield a concentration of 500ug/ml
• Expiration Date: 12 months from date of receipt.

IHC analysis of CD8 alpha using anti-CD8 alpha antibody. CD8 alpha was detected in a paraffin-embedded section of human tonsil tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml rabbit anti-CD8 alpha Antibody overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.

Western blot analysis of CD 8 alpha using anti-CD 8 alpha 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 40 ug of sample under reducing conditions. Lane 1: JURKAT Whole Cell Lysate, Lane 2: RAJI Whole Cell Lysate, Lane 3: HL-60 Whole Cell Lysate, Lane 4: A549 Whole Cell Lysate, Lane 5: COLO320 Whole Cell Lysate, Lane 6: HELA Whole Cell Lysate. 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-CD 8 alpha 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 CD 8 alpha at approximately 55 kDa. The expected band size for CD 8 alpha is at 26 kDa.