Anti-CD55 Antibody (monoclonal, 9F3A1)

• Catalog Number: orb1145782
• Category: Antibodies
• Description: Anti-CD55 Antibody (monoclonal, 9F3A1). Tested in IF, IHC, ICC, WB applications. This antibody reacts with Human.
• Species/Host: Mouse
• Clonality: Monoclonal
• Clone Number: 9F3A1
• Tested applications: ICC, IF, IHC, WB
• Reactivity: Human
• Isotype: Mouse IgG2b
• Immunogen: E.coli-derived human CD55 recombinant protein (Position: D35-K347). Human CD55 shares 49.1% amino acid (aa) sequence identity with mouse CD55.
• 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: 70-75 kDa
• UniProt ID: P08174
• 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: ATP dependent helicase RENT1 antibody; ATP-depende
• Note: For research use only
• Application notes: Western blot, 0.25-0.5 μg/ml, Human Immunohistochemistry(Paraffin-embedded Section), 2-5 μg/ml, Human Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human. Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml
• Expiration Date: 12 months from date of receipt.

IF analysis of CD55 using anti-CD55 antibody. CD55 was detected in an immunocytochemical section of SiHa cells. 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 mouse anti-CD55 Antibody 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.

IHC analysis of CD55 using anti-CD55 antibody. CD55 was detected in a paraffin-embedded section of human placenta 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 mouse anti-CD55 Antibody overnight at 4°C. Peroxidase Conjugated Goat Anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Mouse IgG Super Vision Assay Kit with DAB as the chromogen.

Western blot analysis of CD55 using anti-CD55 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 placenta tissue lysates, Lane 2: human Hela whole cell lysates, Lane 3: human PC-3 whole cell lysates, Lane 4: human Caco-2 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 mouse anti-CD55 antigen affinity purified monoclonal 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-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 with Tanon 5200 system. A specific band was detected for CD55 at approximately 70-75 kDa. The expected band size for CD55 is at 41 kDa.