Anti-CD30/TNFRSF8 Antibody

• Catalog Number: orb669134
• Category: Antibodies
• Description: Anti-CD30/TNFRSF8 Antibody. Tested in IHC, WB applications. This antibody reacts with Human, Mouse.
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: IHC, WB
• Reactivity: Human, Mouse
• Isotype: Rabbit IgG
• Immunogen: A synthetic peptide corresponding to a sequence at the N-terminus of human CD30/TNFRSF8.
• 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: P28908
• 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: Tumor necrosis factor receptor superfamily member
• Note: For research use only
• Application notes: Western blot, 0.5-1μg/ml, Mouse Immunohistochemistry (Paraffin-embedded Section), 2-5μg/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 CD30/TNFRSF8 using anti-CD30/TNFRSF8 antibody. CD30/TNFRSF8 was detected in paraffin-embedded section of human endometrial carcinoma 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 2 µg/ml rabbit anti-CD30/TNFRSF8 Antibody overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Western blot analysis of CD30/TNFRSF8 using anti-CD30/TNFRSF8 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: mouse RAW264.7 whole cell lysates, Lane 2: mouse ANA-1 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-CD30/TNFRSF8 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 CD30/TNFRSF8 at approximately 120 KD. The expected band size for CD30/TNFRSF8 is at 64 KD.