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Western Blot (WB) (Figure 1. Western blot analysis of ARF6 using anti-ARF6 antibody (MBS1753383).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 30ug of sample under reducing conditions.Lane 1: human Hela whole cell lysatesLane 2: human PC-3 whole cell lysatesLane 3: human U20S whole cell lysatesLane 4: rat liver tissue lysatesLane 5: mouse lung tissue lysatesLane 6: mouse liver tissue 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 rabbit anti-ARF6 antigen affinity purified polyclonal antibody (Catalog # MBS1753383) at 0. 5 μg/mL overnight at 4 degree 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 (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for ARF6 at approximately 20KD. The expected band size for ARF6 is at 20KD. )

Rabbit ARF6 Polyclonal Antibody | anti-ARF6 antibody

Anti-ARF6 Antibody

Reactivity
Human, Mouse, Rat
Applications
Western Blot, Immunocytochemistry, Immunofluorescence, Flow Cytometry, Functional Assay, ELISA
Purity
Immunogen affinity purified.
Synonyms
ARF6; Polyclonal Antibody; Anti-ARF6 Antibody; ADP-ribosylation factor 6; ADP ribosylation factor 6; anti-ARF6 antibody
Ordering
For Research Use Only!
Host
Rabbit
Reactivity
Human, Mouse, Rat
Clonality
Polyclonal
Isotype
Rabbit IgG
Specificity
Rabbit IgG polyclonal antibody for ARF6 detection.
Purity/Purification
Immunogen affinity purified.
Form/Format
Lyophilized. Each vial contains 4mg Trehalose, 0.9mg NaCl and 0.2mg Na2HPO4.
Applicable Applications for anti-ARF6 antibody
Western Blot (WB), Immunocytochemistry (ICC), Immunofluorescence (IF), Flow Cytometry (FC/FACS/FCM), Direct ELISA (EIA)
Application Notes
WB: 0.25-0.5ug/ml|Human, Mouse, Rat|
ICC/IF: 5ug/ml|Human|
FC/FACS/FCM: 1-3ug/1x106 cells|Human, Rat|
Direct ELISA: 0.1-0.5ug/ml|Human|
Immunogen
E Coli-derived human ARF6 recombinant protein (Position: R75-S175).
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Recommended Detection Systems
Recommended Detection Systems
Preparation and Storage
Store at -20 degree C for one year. After reconstitution, at 4 degree C for one month. It can also be aliquotted and stored frozen at -20 degree C for a longer time. Avoid repeated freezing and thawing.

Western Blot (WB)

(Figure 1. Western blot analysis of ARF6 using anti-ARF6 antibody (MBS1753383).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 30ug of sample under reducing conditions.Lane 1: human Hela whole cell lysatesLane 2: human PC-3 whole cell lysatesLane 3: human U20S whole cell lysatesLane 4: rat liver tissue lysatesLane 5: mouse lung tissue lysatesLane 6: mouse liver tissue 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 rabbit anti-ARF6 antigen affinity purified polyclonal antibody (Catalog # MBS1753383) at 0. 5 μg/mL overnight at 4 degree 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 (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for ARF6 at approximately 20KD. The expected band size for ARF6 is at 20KD. )

Western Blot (WB) (Figure 1. Western blot analysis of ARF6 using anti-ARF6 antibody (MBS1753383).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 30ug of sample under reducing conditions.Lane 1: human Hela whole cell lysatesLane 2: human PC-3 whole cell lysatesLane 3: human U20S whole cell lysatesLane 4: rat liver tissue lysatesLane 5: mouse lung tissue lysatesLane 6: mouse liver tissue 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 rabbit anti-ARF6 antigen affinity purified polyclonal antibody (Catalog # MBS1753383) at 0. 5 μg/mL overnight at 4 degree 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 (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for ARF6 at approximately 20KD. The expected band size for ARF6 is at 20KD. )

Flow Cytometry (FC/FACS)

(Figure 2. Flow Cytometry analysis of PC-3 cells using anti-ARF6 antibody (MBS1753383).Overlay histogram showing PC-3 cells stained with MBS1753383 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ARF6 Antibody (MBS1753383, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )

Flow Cytometry (FC/FACS) (Figure 2. Flow Cytometry analysis of PC-3 cells using anti-ARF6 antibody (MBS1753383).Overlay histogram showing PC-3 cells stained with MBS1753383 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ARF6 Antibody (MBS1753383, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )

Flow Cytometry (FC/FACS)

(Figure 3. Flow Cytometry analysis of RH35 cells using anti-ARF6 antibody (MBS1753383).Overlay histogram showing RH35 cells stained with MBS1753383 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ARF6 Antibody (MBS1753383, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )

Flow Cytometry (FC/FACS) (Figure 3. Flow Cytometry analysis of RH35 cells using anti-ARF6 antibody (MBS1753383).Overlay histogram showing RH35 cells stained with MBS1753383 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ARF6 Antibody (MBS1753383, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )

Immunofluorescence (IF)

(Figure 4. IF analysis of ARF6 using anti- ARF6 antibody (MBS1753383).ARF6 was detected in immunocytochemical section of A431 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (MBS176582) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti- ARF6 Antibody (MBS1753383) overnight at 4 degree C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used. )

Immunofluorescence (IF) (Figure 4. IF analysis of ARF6 using anti- ARF6 antibody (MBS1753383).ARF6 was detected in immunocytochemical section of A431 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (MBS176582) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti- ARF6 Antibody (MBS1753383) overnight at 4 degree C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used. )
Related Product Information for anti-ARF6 antibody
ADP-ribosylation factor 6 (ARF6) is a member of the human ARF gene family, which is part of the RAS superfamily. The ARF genes encode small guanine nucleotide-binding proteins that stimulate the ADP-ribosyltransferase activity of cholera toxin and play a role in vesicular trafficking and as activators of phospholipase D. The product of this gene is localized to the plasma membrane, and regulates vesicular trafficking, remodelling of membrane lipids, and signaling pathways that lead to actin remodeling. A pseudogene of this gene is located on chromosome 7.
References
1. "Entrez Gene: ARF6 ADP-ribosylation factor 6".
2. Prigent M, Dubois T, Raposo G, Derrien V, Tenza D, Rossé C, Camonis J, Chavrier P (December 2003). "ARF6 controls post-endocytic recycling through its downstream exocyst complex effector". J. Cell Biol. 163(5): 1111-21.
3. Peru y Colón de Portugal RL, Acevedo SF, Rodan AR, Chang LY, Eaton BA, Rothenfluh A (December 2012). "Adult neuronal Arf6 controls ethanol-induced behavior with Arfaptin downstream of Rac1 and RhoGAP18B". J. Neurosci. 32 (49): 17706-13.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
382
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
20,082 Da
NCBI Official Full Name
ADP-ribosylation factor 6
NCBI Official Synonym Full Names
ADP-ribosylation factor 6
NCBI Official Symbol
ARF6
NCBI Protein Information
ADP-ribosylation factor 6
UniProt Protein Name
ADP-ribosylation factor 6
Protein Family
UniProt Gene Name
ARF6
UniProt Entry Name
ARF6_HUMAN

NCBI Description

This gene encodes a member of the human ARF gene family, which is part of the RAS superfamily. The ARF genes encode small guanine nucleotide-binding proteins that stimulate the ADP-ribosyltransferase activity of cholera toxin and play a role in vesicular trafficking and as activators of phospholipase D. The product of this gene is localized to the plasma membrane, and regulates vesicular trafficking, remodelling of membrane lipids, and signaling pathways that lead to actin remodeling. A pseudogene of this gene is located on chromosome 7. [provided by RefSeq, Jul 2008]

Uniprot Description

ARF6: GTP-binding protein involved in protein trafficking; regulates endocytic recycling and cytoskeleton remodeling. May modulate vesicle budding and uncoating within the Golgi apparatus. Functions as an allosteric activator of the cholera toxin catalytic subunit, an ADP-ribosyltransferase. Involved in the regulation of dendritic spine development. Contributes to the regulation of dendritic branching and filopodia extension. Interacts with ARHGAP21, ASAP2, HERC1, PIP5K1C and UACA. Interacts with NCS1/FREQ at the plasma membrane. Interacts with RAB11FIP3 and RAB11FIP4. Interacts with USP6 (via Rab-GAP TBC domain). Interacts with ECM29. Interacts with TBC1D24. Belongs to the small GTPase superfamily. Arf family.

Protein type: G protein, monomeric; Motility/polarity/chemotaxis; G protein, monomeric, ARF

Chromosomal Location of Human Ortholog: 14q21.3

Cellular Component: Golgi apparatus; ruffle; filopodium membrane; focal adhesion; membrane; endocytic vesicle; recycling endosome membrane; early endosome; plasma membrane; cell cortex; midbody; endosome; cleavage furrow

Molecular Function: GTPase activity; protein binding; GTP binding; thioesterase binding

Biological Process: myeloid cell apoptosis; metabolic process; regulation of filopodium formation; liver development; cell cycle; vesicle-mediated transport; regulation of Rac protein signal transduction; protein transport; positive regulation of actin filament polymerization; cell division; small GTPase mediated signal transduction; ruffle organization and biogenesis; negative regulation of receptor-mediated endocytosis; cortical actin cytoskeleton organization and biogenesis; cell adhesion; cell motility

Research Articles on ARF6

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Product Notes

The ARF6 arf6 (Catalog #AAA1753383) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-ARF6 Antibody reacts with Human, Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's ARF6 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunocytochemistry (ICC), Immunofluorescence (IF), Flow Cytometry (FC/FACS/FCM), Direct ELISA (EIA). WB: 0.25-0.5ug/ml|Human, Mouse, Rat| ICC/IF: 5ug/ml|Human| FC/FACS/FCM: 1-3ug/1x106 cells|Human, Rat| Direct ELISA: 0.1-0.5ug/ml|Human|. Researchers should empirically determine the suitability of the ARF6 arf6 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "ARF6, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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