Anti-NUP133 Antibody

• Catalog Number: orb1743784
• Category: Antibodies
• Description: Anti-NUP133 Antibody. Tested in ELISA, IF, ICC, WB applications. This antibody reacts with Human, Rat.
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, ICC, IF, IP, WB
• Reactivity: Human, Mouse, Rat
• Isotype: Rabbit IgG
• Immunogen: E.coli-derived human NUP133 recombinant protein (Position: Q228-I1156).
• 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: 129 kDa
• UniProt ID: Q8WUM0
• 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: Cytoskeleton-associated protein 5; Colonic and hep
• Note: For research use only
• Application notes: Western blot, 0.25-0.5 μg/ml, Human, Mouse, Rat Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human, Rat Immunoprecipitation, 0.5-2 μg/ml, Human ELISA, 0.1-0.5 μg/ml, -. Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml
• Expiration Date: 12 months from date of receipt.

IF analysis of NUP133 using anti-NUP133 antibody and anti-Beta Tubulin antibody. NUP133 was detected in immunocytochemical section of A549 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-NUP133 Antibody and mouse anti-Beta Tubulin antibody overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG and Cy3 Conjugated 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.

IF analysis of NUP133 using anti-NUP133 antibody and anti-Beta Tubulin antibody. NUP133 was detected in immunocytochemical section of C6 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-NUP133 Antibody and mouse anti-Beta Tubulin antibody overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG and DyLight®488 Conjugated 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 NUP133 in Hela whole cell lysate. Western blot analysis of NUP133 using anti-NUP133 antibody. Lane 1: Hela whole cell lysates (30 ug) Lane 2: Rabbit control IgG instead of anti-NUP133 antibody in Hela whole cell lysate. Lane 3: anti-NUP133 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-NUP133 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 NUP133 at approximately 129 kDa. The expected band size for NUP133 is at 129 kDa.

Western blot analysis of NUP133 using anti-NUP133 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 K562 whole cell lysates, Lane 2: human A549 whole cell lysates, Lane 3: human MCF-7 whole cell lysates, Lane 4: rat C6 whole cell lysates, Lane 5: rat NRK whole cell lysates, Lane 6: mouse Neuro-2a whole cell lysates, Lane 7: mouse NIH/3T3 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 rabbit anti-NUP133 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 NUP133 at approximately 129 kDa. The expected band size for NUP133 is at 129 kDa.