Anti-SAE2/UBA2 Antibody (monoclonal, 5H11)

• Catalog Number: orb570311
• Category: Antibodies
• Description: Anti-SAE2/UBA2 Antibody (monoclonal, 5H11). Tested in Flow Cytometry, WB applications. This antibody reacts with Human, Mouse, Rat.
• Species/Host: Mouse
• Clonality: Monoclonal
• Clone Number: 5H11
• Tested applications: FC, WB
• Reactivity: Human, Mouse, Rat
• Isotype: Mouse IgG2b
• Immunogen: E. coli-derived human SAE2/UBA2 recombinant protein (Position: E449-K564).
• Antibody Type: Primary Antibody
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 90 kDa
• UniProt ID: Q9UBT2
• 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, 0.1-0.5μg/ml, Human, Mouse, Rat Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human. Add 0.2ml of distilled water will yield a concentration of 500μg/ml
• Expiration Date: 12 months from date of receipt.

Flow Cytometry analysis of A431 cells using anti-UBA2 antibody. Overlay histogram showing A431 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-UBA2 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 UBA2 using anti-UBA2 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 K562 whole cell lysates Lane 2: human Raji whole cell lysates Lane 3: human THP-1 whole cell lysates Lane 4: human SW579 whole cell lysates Lane 5: human HepG2 whole cell lysates Lane 6: human CCRF-CEM whole cell lysates Lane 7: rat PC-12 whole cell lysates Lane 8: mouse RAW246.7 whole cell lysates After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-UBA2 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 UBA2 at approximately 90KD. The expected band size for UBA2 is at 71KD.