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Products:Signal Transduction >> Protein Trafficking >> Vesicle Transport >> Regulation
Overview
Product name
Anti-ARF6 antibody
See all ARF6 products (6) ...
Description
Rabbit polyclonal to ARF6
Tested applications
ICC/IF, IHC-P, WBmore details
Cross reactivity
Reacts with
Mouse, Rat, Human
Predicted to work with
Chicken, Cow, Pig, Xenopus laevis
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 150 to the C-terminus ARF6.
(Peptide available as ab86625.)
Positive control
This antibody gave a positive signal in the following whole cell lysates: HeLa whole cell, HEK293 whole cell, HepG2 whole cell, Human Liver Tissue
Properties
Form
Liquid
Storage instructions
Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage buffer
Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
Research areas
Signal Transduction >> Protein Trafficking >> Vesicle Transport >> Regulation
Applications
Show applications key
Our Abpromise guarantee covers the use of ab77581 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
ICC/IF: Use a concentration of 1 µg/ml
IHC-P: Use a concentration of 5 µg/mlPerform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB: Use a concentration of 1 µg/mlDetects a band of approximately 18 kDa (predicted molecular weight: 20 kDa).
Target
Relevance
ARF6 (ADP ribosylation factor 6) is a member of the human ARF gene family. These genes encode small guanine nucleotide binding proteins that stimulate the ADP ribosyltransferase activity of cholera toxin and play a role in vesicular trafficking during cellular differentiation and as activators of phospholipase D. The gene products include 6 ARF proteins and 11 ARF like proteins and constitute 1 family of the RAS superfamily. The ARF proteins are categorized as class I (ARF1, ARF2,and ARF3), class II (ARF4 and ARF5) and class III (ARF6) and members of each class share a common gene organization. ARF6 is a broadly expressed, differentially regulated GTPase that is present in cytoplasm and on both cell surface and intracellular membranes.
Cellular localization
Golgi apparatus. Cell membrane; Lipid-anchor. Endosome membrane; Lipid-anchor.
Alternative names
- antibody
- Small GTPase antibody
- ADP ribosylation factor 6 antibody
see all
Database links
- Entrez Gene: 428927 Chicken
- Entrez Gene: 382 Human
- Entrez Gene: 11845 Mouse
- Entrez Gene: 79121 Rat
- Entrez Gene: 399300 Xenopus laevis
- Omim: 600464 Human
- SwissProt: P26990 Chicken
- SwissProt: P62330 Human
see all
Anti-ARF6 antibody images:
Western blot - ARF6 antibody (ab77581)
All lanes : Anti-ARF6 antibody (ab77581) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 4 : Liver (Human) Tissue Lysate - adult normal tissue (ab29889)
Lysates/proteins at 10 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 20 kDa
Observed band size : 18 kDa (why is the actual band size different from the predicted?)
We hypothesize that the 18 kDa band represents the mature form of ARF6.
Immunocytochemistry/ Immunofluorescence - ARF6 antibody (ab77581)
ICC/IF image of ab77581 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab77581, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 100% methanol fixed (5 min) HeLa cells at 1µg/ml.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - ARF6 antibody (ab77581)
IHC image of ARF6 staining in human liver carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab77581, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
Western blot - ARF6 antibody (ab77581)
All lanes : Anti-ARF6 antibody (ab77581) at 1 µg/ml
Lane 1 : Lysates prepared from Mouse liver tissue.
Lane 2 : Lysates prepared from Mouse lung tissue.
Lane 3 : Lysates prepared from Rat liver tissue.
Lane 4 : Lysates prepared from Rat lung tissue.
Lysates/proteins at 20 µg per lane.
Secondary
HRP-conjugated Pig polyclonal to rabbit IgG at 1/5000 dilution
Performed under reducing conditions.
Predicted band size : 20 kDa
Observed band size : 21 kDa (why is the actual band size different from the predicted?)
Exposure time : 2 minutes
This image is a courtesy of Antibody Solutions Ltd
References for Anti-ARF6 antibody (ab77581)
ab77581 has not yet been referenced specifically in any publications.
Publishing research using ab77581? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"