Recombinant Anti-ARF1 + ARF3 antibody [EP442Y] (ab32524)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP442Y] to ARF1 + ARF3
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-ARF1 + ARF3 antibody [EP442Y]
See all ARF1+ARF3 primary antibodies -
Description
Rabbit monoclonal [EP442Y] to ARF1 + ARF3 -
Host species
Rabbit -
Specificity
The immunogen used for this product shares 100% homology with ARF3, ARF4 and ARF5. Cross-reactivity with the latter two protein have not been confirmed experimentally. -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details
Unsuitable for: IP -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide within Human ARF1 aa 50-150. The exact sequence is proprietary.
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Positive control
- HEK-293 cell lysates and human breast carcinoma tissue.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP442Y -
Isotype
IgG -
Research areas
Associated products
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Assay kits
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab32524 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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Flow Cyt (Intra) |
1/80.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB |
1/2000. Detects a band of approximately 20 kDa (predicted molecular weight: 21 kDa).
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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ICC/IF |
1/500.
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Notes |
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Flow Cyt (Intra)
1/80. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/2000. Detects a band of approximately 20 kDa (predicted molecular weight: 21 kDa). |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
ICC/IF
1/500. |
Target
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Cellular localization
ARF1: Golgi apparatus. Cytoplasm > perinuclear region. ARF3: Golgi apparatus. Cytoplasm > perinuclear region. -
Database links
- Entrez Gene: 375 Human
- Entrez Gene: 377 Human
- Entrez Gene: 11840 Mouse
- Entrez Gene: 11842 Mouse
- Entrez Gene: 140940 Rat
- Entrez Gene: 64310 Rat
- Omim: 103180 Human
- Omim: 103190 Human
see all
Images
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Flow cytometry analysis of 4% paraformaldehyde fixed 90% methanol permeabilised MCF7 (human breast adenocarcinoma epithelial cell) cells using ab32524 at 1/500 dilution (red) and secondary Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution. The isotype control was Rabbit monoclonal IgG (ab172730) (Black) and the control was cell swithout incubation with primary antibody and secondary antibody (Blue).
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Anti-ARF1 + ARF3 antibody [EP442Y] (ab32524) at 1/1000 dilution + MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 21 kDa
Observed band size: 20 kDa why is the actual band size different from the predicted?Blocking and diluting buffer: 5% NFDM/TBST. Lysate was freshly made and used for Western blotting immediately to minimize protein degradation.
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All lanes : Anti-ARF1 + ARF3 antibody [EP442Y] (ab32524) at 1/1000 dilution
Lane 1 : SiHa (human cervical epithelial cell) whole cell lysate
Lane 2 : T-47D (human ductal breast epithelial tumor epithelial cell) whole cell lysate
Lysates/proteins at 50 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 21 kDa
Observed band size: 20 kDa why is the actual band size different from the predicted?
Exposure time: 180 seconds
Blocking and diluting buffer: 5% NFDM/TBST -
Immunohistochemistry analysis of paraffin-embedded rat liver tissue sections using ab32524 at 1/2000 dilution and ready to use secondary LeicaDS9800 (Bond™ Polymer Refine Detection). Counterstained with Hematoxylin. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins. Positive staining on rat liver. The section was incubated with ab32524 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument
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Immunohistochemistry analysis of paraffin-embedded mouse liver tissue sections using ab32524 at 1/2000 dilution and ready to use secondary LeicaDS9800 (Bond™ Polymer Refine Detection). Counterstained with Hematoxylin. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins. Positive staining on mouse liver. The section was incubated with ab32524 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument
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All lanes : Anti-ARF1 + ARF3 antibody [EP442Y] (ab32524) at 1/20000 dilution
Lane 1 : Full length human ARF-1 recombinant protein
Lane 2 : Full length human ARF-3 recombinant protein
Lysates/proteins at 0.015 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 21 kDa
Observed band size: 23 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking and diluting buffer: 5% NFDM/TBST
Human ARF-1 recombinant protein, (Cat#:ab123538 )
Human ARF-3 recombinant protein, (Cat#:ab113581 ) -
Immunofluorescence staining of MCF7 cells with purified ab32524 working dilution of 1/500, counter-stained with DAPI. The secondary antibody was an Alexa Fluor® 488 conjugated goat anti-rabbit (ab150077), used at a dilution of 1/1000. The cells were fixed in 100% methanol. The negative control is shown in bottom right hand panel - for the negative control, PBS was used instead of the primary antibody.
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Immunohistochemical analysis of paraffin-embedded human breast carcinoma using ab32524 at 1/250 dilution.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (7)
ab32524 has been referenced in 7 publications.
- Singh V et al. Cell-matrix adhesion controls Golgi organization and function through Arf1 activation in anchorage-dependent cells. J Cell Sci 131:N/A (2018). PubMed: 30054383
- Schultz ML et al. CLN3 deficient cells display defects in the ARF1-Cdc42 pathway and actin-dependent events. PLoS One 9:e96647 (2014). WB ; Mouse . PubMed: 24792215
- Covington JD et al. Skeletal muscle perilipin 3 and coatomer proteins are increased following exercise and are associated with fat oxidation. PLoS One 9:e91675 (2014). WB ; Human . PubMed: 24632837
- Ohashi Y et al. AMF-26, a Novel Inhibitor of the Golgi System, Targeting ADP-ribosylation Factor 1 (Arf1) with Potential for Cancer Therapy. J Biol Chem 287:3885-97 (2012). PubMed: 22158626
- Arita M et al. Phosphatidylinositol 4-kinase III beta is a target of enviroxime-like compounds for antipoliovirus activity. J Virol 85:2364-72 (2011). WB ; Human . PubMed: 21177810
- Anthonio EA et al. Small G proteins in peroxisome biogenesis: the potential involvement of ADP-ribosylation factor 6. BMC Cell Biol 10:58 (2009). PubMed: 19686593
- Luo W et al. p24A, a type I transmembrane protein, controls ARF1-dependent resensitization of protease-activated receptor-2 by influence on receptor trafficking. J Biol Chem 282:30246-55 (2007). PubMed: 17693410