Recombinant Anti-NXF1 antibody [EPR8009] (ab129160)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR8009] to NXF1
- Suitable for: Flow Cyt (Intra), WB, IP, ICC/IF
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-NXF1 antibody [EPR8009]
See all NXF1 primary antibodies -
Description
Rabbit monoclonal [EPR8009] to NXF1 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IP, ICC/IFmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human NXF1. The exact sequence is proprietary.
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Positive control
- WB: HeLa, 293T, K-562 and HepG2 lysates. ICC/IF: HeLa cells. Flow Cyt (intra): HepG2 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.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR8009 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Positive Controls
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab129160 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) |
1/20.
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WB | (1) |
1/1000 - 1/10000. Detects a band of approximately 70 kDa (predicted molecular weight: 70 kDa).
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IP | (1) |
1/10 - 1/100.
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ICC/IF |
1/200 - 1/500.
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Notes |
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Flow Cyt (Intra)
1/20. |
WB
1/1000 - 1/10000. Detects a band of approximately 70 kDa (predicted molecular weight: 70 kDa). |
IP
1/10 - 1/100. |
ICC/IF
1/200 - 1/500. |
Target
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Function
Involved in the nuclear export of mRNA species bearing retroviral constitutive transport elements (CTE) and in the export of mRNA from the nucleus to the cytoplasm. The NXF1-NXT1 heterodimer is involved in the export of HSP70 mRNA in conjunction with THOC4 and THOC5. -
Tissue specificity
Expressed ubiquitously. -
Sequence similarities
Belongs to the NXF family.
Contains 4 LRR (leucine-rich) repeats.
Contains 1 NTF2 domain.
Contains 1 RRM (RNA recognition motif) domain.
Contains 1 TAP-C domain. -
Domain
The minimal CTE binding domain consists of an RNP-type RNA binding domain (RBD) and leucine-rich repeats.
The nucleoporin binding domain consists of a NTF2 domain (also called NTF2-like domain) and a TAP-C domain (also called UBA-like domain). It has 2 nucleoporin-FG-repeats binding sites (one in the NTF2 and the other in the TAP-C domain) which contribute to nucleoporin association and act synergistically to export cellular mRNAs.
The NTF2 domain is functional only in the presence of NXT1 and is essential for the export of mRNA from the nucleus.
The TAP-C domain mediates direct interactions with nucleoporin-FG-repeats and is necessary and sufficient for localization of NXF1 to the nuclear rim. The conserved loop 594-NWD-596 of the TAP-C domain has a critical role in the interaction with nucleoporins.
The leucine-rich repeats are essential for the export of mRNA from the nucleus.
The RNA-binding domain is a non-canonical RNP-type domain. -
Cellular localization
Nucleus > nucleoplasm. Nucleus speckle. Cytoplasm. Localized predominantly in the nucleoplasm and at both the nucleoplasmic and cytoplasmic faces of the nuclear pore complex. Shuttles between the nucleus and the cytoplasm. Travels to the cytoplasm as part of the exon junction complex (EJC) bound to mRNA. - Information by UniProt
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Database links
- Entrez Gene: 10482 Human
- Omim: 602647 Human
- SwissProt: Q9UBU9 Human
- Unigene: 523739 Human
- Unigene: 601546 Human
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Alternative names
- DKFZp667O0311 antibody
- DmNXF1 antibody
- MEX67 antibody
see all
Images
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Anti-NXF1 antibody [EPR8009] (ab129160) at 1/2000 dilution (Purified) + K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysates at 15 µg
Secondary
Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 70 kDa
Observed band size: 70 kDa -
Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling NXF1 with purified ab129160 at 1/200 dilution (0.62 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. 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 (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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Intracellular Flow Cytometry analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling NXF1 with purified ab129160 at 1/20 dilution (5 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 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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All lanes : Anti-NXF1 antibody [EPR8009] (ab129160) at 1/1000 dilution
Lane 1 : HeLa cell lysate
Lane 2 : 293T cell lysate
Lane 3 : K562 cell lysate
Lane 4 : HepG2 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat anti-rabbit HRP at 1/2000 dilution
Predicted band size: 70 kDa -
ab129160 at 1/250 dilution staining NXF1 in HepG2 cells by immunofluorescence.
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ab129160 at 1/20 immunoprecipitating NXF1 in HepG2 (human hepatocellular carcinoma) whole cell lysate observed at 70 KDa (Lanes 1 and 2).
Lane 1 (input): HepG2 whole cell lysate, 10μg
Lane 2 (+): ab129160 + HepG2 whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab129160 in HepG2 whole cell lysateFor western blotting, ab129160 at 1/500 dilution and ab131366 VeriBlot for IP (HRP) at 1/1000 were used.
Blocking and Diluting buffer and concentration: 5% NFDM/TBST.
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ab129160 at 1/20 immunoprecipitating NXF1 in HeLa (human cervix adenocarcinoma) whole cell lysate observed at 70 KDa (Lanes 1 and 2).
Lane 1 (input): HeLa whole cell lysate, 10μg
Lane 2 (+): ab129160 + HeLa whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab129160 in HeLa whole cell lysateFor western blotting, ab129160 at 1/500 dilution and ab131366 VeriBlot for IP (HRP) at 1/1000 were used.
Blocking and Diluting buffer and concentration: 5% NFDM/TBST.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (4)
ab129160 has been referenced in 4 publications.
- He Y et al. T-cell receptor (TCR) signaling promotes the assembly of RanBP2/RanGAP1-SUMO1/Ubc9 nuclear pore subcomplex via PKC-θ-mediated phosphorylation of RanGAP1. Elife 10:N/A (2021). PubMed: 34110283
- Fáberová V et al. Super-Resolution Localisation of Nuclear PI(4)P and Identification of Its Interacting Proteome. Cells 9:N/A (2020). PubMed: 32403279
- Ben-Yishay R et al. Imaging within single NPCs reveals NXF1's role in mRNA export on the cytoplasmic side of the pore. J Cell Biol 218:2962-2981 (2019). PubMed: 31375530
- Roundtree IA et al. YTHDC1 mediates nuclear export of N6-methyladenosine methylated mRNAs. Elife 6:N/A (2017). PubMed: 28984244