Angiogenin/ANG Antibody

SKU: orb389386

Description

Anti-Angiogenin/ANG Antibody. Tested in ELISA, Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human.

Images & Validation

• Tested Applications: ELISA, FC, ICC, IF, IHC, WB
• Reactivity: Human
• Application Notes:
  Western blot, 0.1-0.5μg/ml, Human Immunohistochemistry(Paraffin-embedded Section), 2-5 μg/ml, Human Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human Flow Cytometry(Fixed), 1-3 μg/1x106 cells, Human ELISA(Cap) , 1-5μg/ml, Human, -. Add 0.2ml of distilled water will yield a concentration of 500ug/ml

Key Properties

• Antibody Type: Primary Antibody
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: Rabbit IgG
• Immunogen: E. coli-derived human Angiogenin/ANG recombinant protein (Position: Q25-P147). Human Angiogenin/ANG shares 77.5% amino acid (aa) sequence identity with mouse Angiogenin/ANG.
• Molecular Weight: 17 kDa
• Purification: Immunogen affinity purified.

Storage & Handling

• 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.
• Form/Appearance: Lyophilized
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Disclaimer: For research use only

Alternative Names

Angiogenin; 3.1.27.-; Ribonuclease 5; RNase 5; ANG; RNASE5

Flow Cytometry analysis of HepG2 cells using anti-Angiogenin/ANG antibody. Overlay histogram showing HepG2 cells (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-Angiogenin/ANG 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 (Red line) was also used as a control.

IF analysis of Angiogenin/ANG using anti-Angiogenin/ANG antibody and anti-Beta Tubulin antibody. Angiogenin/ANG 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-Angiogenin/ANG Antibody and mouse anti-Beta Tubulin antibody overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG and FITC 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.

IHC analysis of Angiogenin/ANG using anti-Angiogenin/ANG antibody. Angiogenin/ANG was detected in a paraffin-embedded section of human hepatocellular carcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml rabbit anti-Angiogenin/ANG Antibody overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.

IHC analysis of Angiogenin/ANG using anti-Angiogenin/ANG antibody. Angiogenin/ANG was detected in a paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml rabbit anti-Angiogenin/ANG Antibody overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.

Sandwich ELISA - Recombinant human ANG protein standard curve. Use in combination with reagents from Human ANG ELISA Kit EZ-Set (DIY Antibody Pairs).

Western blot analysis of Angiogenin/ANG using anti-Angiogenin/ANG 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 hepatocellular carcinoma tumor tissue (HCCT) 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-Angiogenin/ANG 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 Angiogenin/ANG at approximately 17 kDa. The expected band size for Angiogenin/ANG is at 17 kDa.

Quick Database Links

UniProt

P03950