Anti-AFF4 Antibody (monoclonal, 8G12)

• Catalog Number: orb738417
• Category: Antibodies
• Description: Anti-AFF4 Antibody (monoclonal, 8G12). Tested in Flow Cytometry, WB applications. This antibody reacts with Human.
• Species/Host: Mouse
• Clonality: Monoclonal
• Clone Number: 8G12
• Tested applications: FC, WB
• Reactivity: Human
• Isotype: Mouse IgG2b
• Immunogen: A synthetic peptide corresponding to a sequence at the N-terminus of human AFF4, identical to the related mouse sequence.
• 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: 150 kDa
• UniProt ID: Q9UHB7
• 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: SUMO-activating enzyme subunit 2; Anthracycline-as
• Note: For research use only
• Application notes: Western blot, 0.25-0.5μg/ml, Human Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human. Add 0.2ml of distilled water will yield a concentration of 500ug/ml
• Expiration Date: 12 months from date of receipt.

Flow Cytometry analysis of 293T cells using anti-AFF4 antibody. Overlay histogram showing 293T cells (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-AFF4 Antibody (1 µg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (5-10 µg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1 µg/1x10^6) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Western blot analysis of AFF4 using anti-AFF4 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 50 ug of sample under reducing conditions. Lane 1: human HELA whole cell lysates, Lane 2: human HEPG2 whole cell lysates, Lane 3: human CACO-2 whole cell lysates, Lane 4: human HEK293 whole cell lysates, Lane 5: human MDA-MB-453 whole cell lysates, Lane 6: human PANC-1 whole cell lysates, Lane 7: human SW620 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-AFF4 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 AFF4 at approximately 150 KD. The expected band size for AFF4 is at 150 KD.