ARF1 Recombinant Rabbit Monoclonal Antibody [SD2002]
Catalog# ET1612-12
ARF1 Recombinant Rabbit Monoclonal Antibody [SD2002]
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WB
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IHC-P
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FC
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Human
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Mouse
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Rat
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unconjugated
Overview
Product Name
ARF1 Recombinant Rabbit Monoclonal Antibody [SD2002]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within Human ARF1aa 61-110 / 181.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IHC-P, FC
Molecular Weight
21 kDa
Positive Control
NIH/3T3 cell lysates, human colon carcinoma tissue, human breast carcinoma tissue, Hela.
Conjugation
unconjugated
Clone Number
SD2002
RRID
Product Features
Form
Liquid
Concentration
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:1,000-1:2,000
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FC
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1:50-1:100
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IHC-P
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1:100
Target
Function
ADP-ribosylation factors (ARFs), are small guanine nucleotide-binding proteins that enhance the enzymatic activities of cholera toxin, and constitute one family of the RAS superfamily. ARFs are essential and ubiquitous in eukaryotes, as they are involved in vesicular transport and functioning via phospholipase D activation. ARF proteins play a role in membrane traffic and organelle integrity and are intimately tied to their reversible association with membranes and distinct interactions with membrane phospholipids. ARF1 is regulated by the binding and hydrolysis of GTP. Coatomer, or COPI, is a heptameric protein recruited to membranes by ARF1. Research demonstrates that guanine nucleotide exchange-activated ARF1, when located at the Golgi membrane, recruits and binds cytoplasmic COPI to the membranes.
Background References
1. Schultz ML et al. CLN3 deficient cells display defects in the ARF1-Cdc42 pathway and actin-dependent events. PLoS One 9:e96647 (2014).
2. Jung JJ et al. Secretion of soluble vascular endothelial growth factor receptor 1 (sVEGFR1/sFlt1) requires Arf1, Arf6, and Rab11 GTPases. PLoS One 7:e44572 (2012).
Sequence Similarity
Belongs to the small GTPase superfamily. Arf family.
Post-translational Modification
Demyristoylated by S.flexneri cysteine protease IpaJ which cleaves the peptide bond between N-myristoylated Gly-2 and Asn-3.
Subcellular Location
Golgi apparatus, Cytoplasm, Cell junction , Membrane.
Synonyms
ADP Ribosylation Factor 1 antibody
ADP-ribosylation factor 1 antibody
ARF 1 antibody
ARF1 antibody
ARF1_HUMAN antibody
Images
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☑ Knockdown (KD)
Western blot analysis of ARF1 on different lysates with Rabbit anti-ARF1 antibody (ET1612-12) at 1/2,000 dilution.
Lane 1: HAP1-parental cell lysate
Lane 2: HAP1-ARF1 KD cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 18 kDa
Observed band size: 18 kDa
Exposure time: 180 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1612-12) at 1/2,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Western blot analysis of ARF1 on NIH/3T3 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1612-12, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
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Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-ARF1 antibody (ET1612-12) at 1/100 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-12) at 1/100 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-ARF1 antibody (ET1612-12) at 1/100 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-12) at 1/100 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Flow cytometric analysis of ARF1 was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1612-12, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"