Ran Mouse Monoclonal Antibody

SKU: orb570321

Description

Anti-Ran Antibody (monoclonal, 5D5). Tested in Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.

Research Area

Cancer Biology, Metabolism Research, Neuroscience, Pharmacology & Drug Discovery, Signal Transduction

Images & Validation

• Tested Applications: FC, ICC, IF, WB
• Dilution Range: Western blot, 0.1-0.5μg/ml Immunocytochemistry/Immunofluorescence, 2μg/ml Flow Cytometry (Fixed), 1-3μg/1x10^6 cells
• Reactivity: Human, Mouse, Rat

Key Properties

• Antibody Type: Primary Antibody
• Host: Mouse
• Clonality: Monoclonal
• Isotype: Mouse IgG2b
• Clone No.: 5D5
• Immunogen: E. coli-derived human Ran recombinant protein (Position: A2-L216). Human Ran shares 100% amino acid (aa) sequence identity with both mouse and rat Ran.
• Target: GTP-binding nuclear protein Ran
• Molecular Weight: 24 kDa
• Purification: Immunogen affinity purified.
• Conjugation: Unconjugated

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
• Buffer/Preservatives: Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
• Concentration: 500 µg/ml
• Expiration Date: 12 months from date of receipt.
• Disclaimer: For research use only

Alternative Names

ARA24; Gsp1; GTPase Ran; RAN; Ras like protein TC4; Ras related nuclear protein; TC4

Flow Cytometry analysis of PC-3 cells using anti-Ran antibody. Overlay histogram showing PC-3 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 mouse anti-Ran Antibody (1 µg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (5-10 µg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse 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 Ran using anti-Ran antibody. Ran was detected in immunocytochemical section of U20S 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 2 µg/mL mouse anti-Ran Antibody overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Western blot analysis of Ran using anti-Ran 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 50 ug of sample under reducing conditions. Lane 1: human HL-60 whole cell lysates, Lane 2: human T-47D whole cell lysates, Lane 3: human A549 whole cell lysates, Lane 4: human U2OS whole cell lysates, Lane 5: human THP-1 whole cell lysates, Lane 6: human HepG2 whole cell lysates, Lane 7: human PANC-1 whole cell lysates, Lane 8: human SW620 whole cell lysates, After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-Ran antigen affinity purified monoclonal 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-mouse IgG-HRP secondary antibody at a dilution of 1:10000 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 Ran at approximately 24KD. The expected band size for Ran is at 24KD.

Western blot analysis of Ran using anti-Ran 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 50 ug of sample under reducing conditions. Lane 1: rat testis tissue lysates, Lane 2: mouse lung tissue lysates, Lane 3: mouse kidney tissue lysates, Lane 4: mouse testis tissue lysates, Lane 5: mouse Neuro-2a whole cell lysates, After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-Ran antigen affinity purified monoclonal 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-mouse IgG-HRP secondary antibody at a dilution of 1:10000 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 Ran at approximately 24KD. The expected band size for Ran is at 24KD.

Quick Database Links

Gene Symbol

GTP-binding nuclear protein Ran

UniProt

P62826