Anillin/ANLN Antibody

• Catalog Number: orb526983
• Category: Antibodies
• Description: Anti-Anillin/ANLN Antibody. Tested in Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human.
• Clonality: Polyclonal
• Species/Host: Rabbit
• Isotype: Rabbit IgG
• Conjugation: Unconjugated
• Reactivity: Human, Mouse, 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: A synthetic peptide corresponding to a sequence at the C-terminus of human Anillin/ANLN, which shares 96.4% amino acid (aa) sequence identity with both mouse and rat Anillin/ANLN.
• UniProt ID: Q9NQW6
• MW: 150 kDa
• Tested applications: FC, ICC, IF, IP, WB
• Application notes: Western blot, 0.1-0.5μg/ml Immunocytochemistry/Immunofluorescence, 5 μg/ml Immunoprecipitation, 0.5-2 μg/ml, Human Flow Cytometry (Fixed), 1-3μg/1x106 cells. 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: Anillin; ANLN
• Note: For research use only

Flow Cytometry analysis of U251 cells using anti-Anillin/ANLN antibody. Overlay histogram showing U251 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-Anillin/ANLN 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 Anillin/ANLN using anti-Anillin/ANLN antibody and anti-Tubulin Alpha antibody. Anillin/ANLN was detected in immunocytochemical section of U2OS 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-Anillin/ANLN Antibody and mouse anti-Tubulin Alpha antibody overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG and Cy3 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 Anillin/ANLN in U251 whole cell lysate. Western blot analysis of Anillin/ANLN using anti-Anillin/ANLN antibody; Lane 1: U251 whole cell lysates (30 ug); Lane 2: Rabbit control IgG instead of anti-Anillin/ANLN antibody in U251 whole cell lysate; Lane 3: anti-Anillin/ANLN antibody (2 µg) + U251 whole cell lysate (500 µg). After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-Anillin/ANLN 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 Anillin/ANLN at approximately 150 kDa. The expected band size for Anillin/ANLN is at 124 kDa.

Western blot analysis of Anillin/ANLN using anti-Anillin/ANLN antibody. Electrophoresis was performed on a 8% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 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 MCF-7 whole cell lysates, Lane 3: rat PC-12 whole cell lysates, Lane 4: 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-Anillin/ANLN 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 ECL Plus Western Blotting Substrate with Tanon 5200 system. A specific band was detected for Anillin/ANLN at approximately 150 kDa. The expected band size for Anillin/ANLN is at 124 kDa.