SAE2/UBA2 Antibody (monoclonal, 5H11)

• Catalog Number: orb570311
• Category: Antibodies
• Description: SAE2/UBA2 Antibody (monoclonal, 5H11)
• 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).
• Concentration: 0
• Dilution range: Western blot, 0.1-0.5μg/ml, Human, Mouse, Rat Flow Cytometry, 1-3μg/1x106 cells, Human
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 90 kDa
• UniProt ID: Q9UBT2
• Storage: Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
• Alternative names: SUMO-activating enzyme subunit 2; Anthracycline-as
• Note: For research use only
• Application notes: Tested Species: In-house tested species with positive results. By Heat: Boiling the paraffin sections in 10mM citrate buffer, pH6.0, for 20mins is required for the staining of formalin/paraffin sections. Other applications have not been tested. Optimal dilutions should be determined by end users. Add 0.2ml of distilled water will yield a concentration of 500μg/ml.
• Expiration Date: 12 months from date of receipt.

Western blot analysis of UBA2 using anti-UBA2 antibody (orb570311). 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 50ug 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 (Catalog # orb570311) 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 (Catalog # orb90502) with Tanon 5200 system. A specific band was detected for UBA2 at approximately 90KD. The expected band size for UBA2 is at 71KD.

Flow Cytometry analysis of A431 cells using anti-UBA2 antibody (orb570311). Overlay histogram showing A431 cells stained with orb570311 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-UBA2 Antibody (orb570311, 1μg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (5-10μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Flow Cytometry analysis of U20S cells using anti-UBA2 antibody (orb570311). Overlay histogram showing U20S cells stained with orb570311 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-UBA2 Antibody (orb570311, 1μg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (5-10μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.