ARFGAP1 Rabbit Polyclonal Antibody

SKU: orb389390

Description

Anti-ARFGAP1 Antibody. Tested in Flow Cytometry, ICC, IHC, IF, WB applications. This antibody reacts with Human, Mouse, Rat.

Research Area

Cell Biology, Protein Biochemistry, Signal Transduction

Images & Validation

• Tested Applications: FC, ICC, IF, IHC, WB
• Dilution Range: Western blot, 0.1-0.5μg/ml, Human, Mouse, Rat Immunohistochemistry(Paraffin-embedded Section), 2-5 μg/ml, Mouse, Rat Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human Flow Cytometry (Fixed), 1-3 μg/1x10^6 cells, Human
• Reactivity: Human, Mouse, Rat

Key Properties

• Antibody Type: Primary Antibody
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: Rabbit IgG
• Immunogen: E. coli-derived human ARFGAP1 recombinant protein (Position: M1-Y183). Human ARFGAP1 shares 89.1% and 88.5% amino acid (aa) sequence identity with mouse and rat ARFGAP1, respectively.
• Target: ADP-ribosylation factor GTPase-activating protein 1
• Molecular Weight: 45 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: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Expiration Date: 12 months from date of receipt.
• Disclaimer: For research use only

Alternative Names

ADP-ribosylation factor GTPase-activating protein 1; ARF GAP 1; ADP-ribosylation factor 1 GTPase-activating protein; ARF1 GAP; ARF1-ed GTPase-activating protein; ARFGAP1; ARF1GAP

Flow Cytometry analysis of HepG2 cells using anti-ARFGAP1 antibody. Overlay histogram showing HepG2 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-ARFGAP1 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 ARFGAP1 using anti-ARFGAP1 antibody. ARFGAP1 was detected in an immunocytochemical section of CACO-2 cells. 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-ARFGAP1 Antibody overnight at 4°C. DyLight594 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

IHC analysis of ARFGAP1 using anti-ARFGAP1 antibody. ARFGAP1 was detected in a paraffin-embedded section of mouse brain 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-ARFGAP1 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 ARFGAP1 using anti-ARFGAP1 antibody. ARFGAP1 was detected in a paraffin-embedded section of rat brain 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-ARFGAP1 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.

Western blot analysis of ARFGAP1 using anti-ARFGAP1 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 MOLT-4 whole cell lysates, Lane 2: human Caco-2 whole cell lysates, Lane 3: human HepG2 whole cell lysates, Lane 4: rat brain tissue lysates, Lane 5: mouse brain 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-ARFGAP1 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 ARFGAP1 at approximately 45 kDa. The expected band size for ARFGAP1 is at 45 kDa.

Quick Database Links

Gene Symbol

ADP-ribosylation factor GTPase-activating protein 1

UniProt

Q8N6T3