Anti-CD20 Antibody (monoclonal, 4D11)

• Catalog Number: orb1145785
• Category: Antibodies
• Description: Anti-CD20 Antibody (monoclonal, 4D11). Tested in IF, IHC, WB applications. This antibody reacts with Human.
• Clonality: Monoclonal
• Species/Host: Mouse
• Isotype: Mouse IgG2b
• Conjugation: Unconjugated
• Reactivity: Human
• Form/Appearance: Lyophilized
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Immunogen: E.coli-derived human CD20 recombinant protein (Position: M1-D261). Human CD20 shares 75% amino acid (aa) sequence identity with mouse CD20.
• UniProt ID: P11836
• MW: 33 kDa
• Tested applications: IF, IHC, WB
• Application notes: Western blot, 0.25-0.5 μg/ml, Human Immunohistochemistry(Paraffin-embedded Section), 2-5 μg/ml, Human Immunofluorescence, 5 μg/ml, Human. Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml
• Antibody Type: Primary Antibody
• Clone Number: 4D11
• 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: Eukaryotic translation initiation factor 6; eIF-6;
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

IF analysis of CD3E and CD20 using anti-CD3E antibody and anti-CD20 antibody. CD3E and CD20 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 5 µg/mL rabbit anti-CD3E antibody and mouse anti-CD20 antibody overnight at 4°C. DyLight®550 Conjugated Goat Anti-Rabbit IgG, DyLight®488 Conjugated Goat Anti-Mouse IgG was used as secondary antibody at 1:500 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 CD20 using anti-CD20 antibody. CD20 was detected in a paraffin-embedded section of human gastric 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 mouse anti-CD20 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.

IHC analysis of CD20 using anti-CD20 antibody. CD20 was detected in a paraffin-embedded section of human lienal rupture 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-CD20 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 CD20 using anti-CD20 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 Jurkat whole cell lysates, Lane 2: human Raji 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-CD20 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 1:5000 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 CD20 at approximately 33 kDa. The expected band size for CD20 is at 33 kDa.