HIP2/LIG/UBE2K Antibody

• Catalog Number: orb865579
• Category: Antibodies
• Description: HIP2/LIG/UBE2K Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, FC, WB
• Reactivity: Human, Monkey, Mouse, Rat
• Isotype: Rabbit IgG
• Immunogen: E.coli-derived human HIP2/LIG/UBE2K recombinant protein (Position: D33-E195).
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Dilution range: Western blot, 0.1-0.25 μg/ml, Human, Mouse, Rat, Monkey Flow Cytometry, 1-3 μg/1x106 cells, Rat Direct ELISA, 0.1-0.5 μg/ml, Human
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 22 kDa
• UniProt ID: P61086
• Storage: 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 freezing and thawing.
• Note: For research use only
• Application notes: Tested Species: In-house tested species with positive results. Other applications have not been tested. Optimal dilutions should be determined by end users. Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Expiration Date: 12 months from date of receipt.

Western blot analysis of HIP2/LIG/UBE2K using anti-HIP2/LIG/UBE2K antibody (orb865579). 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: monkey COS-7 whole cell lysates, Lane 2: human HEK293 whole cell lysates, Lane 3: rat brain tissue lysates, Lane 4: rat testis tissue lysates, Lane 5: mouse brain tissue lysates, Lane 6: mouse testis tissue 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 rabbit anti-HIP2/LIG/UBE2K antigen affinity purified polyclonal antibody (Catalog # orb865579) at 0.25 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-rabbit 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 (Catalog # orb90503) with Tanon 5200 system. A specific band was detected for HIP2/LIG/UBE2K at approximately 22 kDa. The expected band size for HIP2/LIG/UBE2K is at 22 kDa.

Flow Cytometry analysis of RH35 cells using anti-HIP2/LIG/UBE2K antibody (orb865579). Overlay histogram showing RH35 cells stained with orb865579 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-HIP2/LIG/UBE2K Antibody (orb865579, 1 μg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10 μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.