Recombinant Anti-Rab4 antibody [EPR3043] - Early Endosome Marker (ab109009)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR3043] to Rab4 - Early Endosome Marker
- Suitable for: Flow Cyt (Intra), WB, IP, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Rab4 antibody [EPR3043] - Early Endosome Marker
See all Rab4 primary antibodies -
Description
Rabbit monoclonal [EPR3043] to Rab4 - Early Endosome Marker -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IP, ICC/IFmore details
Unsuitable for: IHC-P -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: MCF7, PC12, Neuro 2a, 293T, SH SY5Y and Human fetal brain lysates; ICC/IF: HeLa cells. Flow Cyt (intra): HeLa cells. IP: MCF7 cells.
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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. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), PBS, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR3043 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab109009 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) |
1/200.
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WB |
1/1000 - 1/10000. Predicted molecular weight: 24 kDa.
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IP |
1/10 - 1/100.
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ICC/IF | (1) |
1/170 - 1/1000.
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Notes |
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Flow Cyt (Intra)
1/200. |
WB
1/1000 - 1/10000. Predicted molecular weight: 24 kDa. |
IP
1/10 - 1/100. |
ICC/IF
1/170 - 1/1000. |
Target
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Function
Protein transport. Probably involved in vesicular traffic. -
Sequence similarities
Belongs to the small GTPase superfamily. Rab family. -
Post-translational
modificationsPhosphorylated by CDK1 kinase during mitosis. -
Cellular localization
Membrane. Cytoplasm. Generally associated with membranes. Cytoplasmic when phosphorylated by CDK1. - Information by UniProt
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Database links
- Entrez Gene: 5867 Human
- Entrez Gene: 19341 Mouse
- Entrez Gene: 25532 Rat
- Omim: 179511 Human
- SwissProt: P20338 Human
- SwissProt: P56371 Mouse
- SwissProt: P05714 Rat
- Unigene: 296169 Human
see all -
Alternative names
- HRES 1 / RAB4 antibody
- Oncogene RAB4 antibody
- Rab 4 antibody
see all
Images
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Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Rab4 with Purified ab109009 at 1:170 dilution (10 µg/ml). Cells were fixed in 100% Methanol. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: Rab4 knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: Human fetal brain lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab109009 observed at 24 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab109009 was shown to specifically react with Rab4 when Rab4 knockout samples were used. Wild-type and Rab4 knockout samples were subjected to SDS-PAGE. ab109009 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/2000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging. -
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution (Purified)
Lane 1 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates
Lane 2 : Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysates
Lane 3 : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysates
Lysates/proteins at 1/15 dilution per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 24 kDa -
ab109009 (purified) at 1:80 dilution (2µg) immunoprecipitating Rab4 in MCF7 whole cell lysate.
Lane 1 (input): MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate 10µg
Lane 2 (+): ab109009 & MCF7 whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab109009 in MCF7 whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST. -
Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Rab4 with Purified ab109009 at 1/200 dilution (1µg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluorr® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) labelling Rab4 with purified ab109009 at 1/250. Cells were fixed with 100% methanol and permeabilized with 0.1% triton X-100. ab150077 Goat anti rabbit IgG (Alexa Fluor® 488) at 1/1000 was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.
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All lanes : Anti-Rab4 antibody [EPR3043] - Early Endosome Marker (ab109009) at 1/1000 dilution
Lane 1 : MCF7 cell lysate
Lane 2 : PC12 cell lysate
Lane 3 : Neuro 2a cell lysate
Lane 4 : 293T cell lysate
Lane 5 : SH SY5Y cell lysate
Lane 6 : Human fetal brain lysate
Lysates/proteins at 10 µg per lane.
Predicted band size: 24 kDa -
ab109009 at 1/500 dilution staining Rab4 in HeLa by Immunofluorescence.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (10)
ab109009 has been referenced in 10 publications.
- Guo Z et al. Activity-dependent PI4P synthesis by PI4KIIIα regulates long-term synaptic potentiation. Cell Rep 38:110452 (2022). PubMed: 35235793
- Chen PM et al. CD38 reduces mitochondrial fitness and cytotoxic T cell response against viral infection in lupus patients by suppressing mitophagy. Sci Adv 8:eabo4271 (2022). PubMed: 35704572
- Yu Z et al. Hepatocyte growth factor-regulated tyrosine kinase substrate is essential for endothelial cell polarity and cerebrovascular stability. Cardiovasc Res 117:533-546 (2021). PubMed: 32044971
- Salvany S et al. Microglial recruitment and mechanisms involved in the disruption of afferent synaptic terminals on spinal cord motor neurons after acute peripheral nerve injury. Glia 69:1216-1240 (2021). PubMed: 33386754
- Rivero-Ríos P et al. Distinct Roles for RAB10 and RAB29 in Pathogenic LRRK2-Mediated Endolysosomal Trafficking Alterations. Cells 9:N/A (2020). PubMed: 32709066
- Gong B et al. Sec14l3 potentiates VEGFR2 signaling to regulate zebrafish vasculogenesis. Nat Commun 10:1606 (2019). PubMed: 30962435
- Rivero-Ríos P et al. The G2019S variant of leucine-rich repeat kinase 2 (LRRK2) alters endolysosomal trafficking by impairing the function of the GTPase RAB8A. J Biol Chem 294:4738-4758 (2019). PubMed: 30709905
- Johnson IRD et al. A Paradigm in Immunochemistry, Revealed by Monoclonal Antibodies to Spatially Distinct Epitopes on Syntenin-1. Int J Mol Sci 20:N/A (2019). PubMed: 31795513
- Xin X et al. Drug-delivering-drug platform-mediated potent protein therapeutics via a non-endo-lysosomal route. Theranostics 8:3474-3489 (2018). PubMed: 30026860
- Niu Y et al. Ablation of SNX6 leads to defects in synaptic function of CA1 pyramidal neurons and spatial memory. Elife 6:N/A (2017). PubMed: 28134614