Anti-C5/C5a Antibody

• Catalog Number: orb215941
• Category: Antibodies
• Description: Anti-C5/C5a Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: IHC, WB
• Reactivity: Rat
• Isotype: Rabbit IgG
• Immunogen: E.coli-derived rat C5a recombinant protein (Position: D1-R77). Rat C5a shares 62% and 82% amino acid (aa) sequences identity with human and mouse C5a, 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: 115 kDa
• UniProt ID: P08650
• 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: Complement C5; C5a anaphylatoxin; C5
• Note: For research use only
• Application notes: Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, RatWestern blot, 0.1-0.5μg/ml, Rat. Add 0.2ml of distilled water will yield a concentration of 500ug/ml
• Expiration Date: 12 months from date of receipt.

IHC analysis of C5A using anti-C5A antibody. C5A was detected in a paraffin-embedded section of rat spleen tissue. Heat mediated antigen retrieval was performed in EDTAbuffer (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-C5A 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 C5A using anti-C5A 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: rat liver 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 rabbit anti-C5A 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 C5A at approximately 115 kDa. The expected band size for C5A is at 115 kDa.

Western blot analysis of C5A using anti-C5A antibody. Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. Lane 1: recombinant rat C5A protein 0.5 ng. 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-C5A 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 C5A at approximately 37 kDa. The expected band size for C5A is at 37 kDa.