DIS3 Antibody

• Catalog Number: orb526991
• Category: Antibodies
• Description: Anti-DIS3 Antibody. Tested in ELISA, Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Rat.
• Clonality: Polyclonal
• Species/Host: Rabbit
• Isotype: Rabbit IgG
• Conjugation: Unconjugated
• Reactivity: Human, Rat
• Form/Appearance: Lyophilized
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Purification: Immunogen affinity purified.
• Immunogen: E. coli-derived human DIS3 recombinant protein (Position: Q62-A304).
• UniProt ID: Q9Y2L1
• MW: 109 kDa
• Tested applications: ELISA, FC, ICC, IF, IP, WB
• Application notes: Western blot, 0.1-0.5μg/ml, Human, Rat Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human Immunofluorescence, 5 μg/ml, Human Immunoprecipitation, 0.5-2 μg/ml, Human Flow Cytometry(Fixed), 1-3 μg/1x106 cells, Human ELISA, 0.1-0.5μg/ml. Add 0.2ml of distilled water will yield a concentration of 500ug/ml
• Cross Reactivity: No cross-reactivity with other proteins.
• Antibody Type: Primary Antibody
• 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: Exosome complex exonuclease RRP44; Protein DIS3 ho
• Note: For research use only

Flow Cytometry analysis of MCF-7 cells using anti-DIS3 antibody. Overlay histogram showing MCF-7 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 rabbit anti-DIS3 Antibody (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 without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

IF analysis of DIS3 using anti-DIS3 antibody and anti-Beta Tubulin antibody. DIS3 was detected in immunocytochemical section of Hela cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 µg/mL rabbit anti-DIS3 Antibody and mouse anti-Beta Tubulin antibody overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG and DyLight®594 Conjugated Goat Anti-Mouse IgG were used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Immunoprecipitating DIS3 in Hela whole cell lysate. Western blot analysis of DIS3 using anti-DIS3 antibody; Lane 1: Hela whole cell lysates (30 ug); Lane 2: Rabbit control IgG instead of anti-DIS3 antibody in Hela whole cell lysate; Lane 3: anti-DIS3 antibody (2 µg) + Hela whole cell lysate (500 µg). After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-DIS3 antigen affinity purified polyclonal antibody at a dilution of 0.5 µg/mL and probed with a goat anti-rabbit IgG-HRP secondary antibody. The signal is developed using ECL Plus Western Blotting Substrate. A specific band was detected for DIS3 at approximately 109 kDa. The expected band size for DIS3 is at 109 kDa.

Western blot analysis of DIS3 using anti-DIS3 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 Hela whole cell lysates, Lane 2: human Raji whole cell lysates, Lane 3: human MCF-7 whole cell lysates, Lane 4: rat 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-DIS3 antigen affinity purified polyclonal 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-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 with Tanon 5200 system. A specific band was detected for DIS3 at approximately 109 kDa. The expected band size for DIS3 is at 109 kDa.