Name: Anti-NXF1 antibody [EPR8009]
Brand: RabMAb
SKU: ab129160
Rating: 5 (2 reviews)

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Western blot - 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: WB, IP, Flow Cyt, ICC/IF
Reacts with: Human

Product name

Anti-NXF1 antibody [EPR8009]
See all NXF1 primary antibodies
Description

Rabbit monoclonal [EPR8009] to NXF1
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Human
IP	Human
WB	Human

See all applications and species data

Immunogen

Synthetic peptide within Human NXF1. The exact sequence is proprietary.
Positive control
- WB: HeLa, 293T, K-562 and HepG2 lysates. ICC/IF: HeLa cells. Flow Cyt: HepG2 cells.
General notes
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

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...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR8009
Isotype

IgG
Research areas

Alternative Versions
- Anti-NXF1 antibody [EPR8009] - BSA and Azide free (ab248319)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Positive Controls
Recombinant Protein
- Recombinant Human NXF1 protein (ab153087)

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.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application	Species
Flow Cyt	Human
ICC/IF	Human
IP	Human
WB	Human

Application	Abreviews	Notes
WB	(1)	1/1000 - 1/10000. Detects a band of approximately 70 kDa (predicted molecular weight: 70 kDa).
IP	(1)	1/10 - 1/100.
Flow Cyt		1/20.
ICC/IF		1/200 - 1/500.

Notes
WB 1/1000 - 1/10000. Detects a band of approximately 70 kDa (predicted molecular weight: 70 kDa).
IP 1/10 - 1/100.
Flow Cyt 1/20.
ICC/IF 1/200 - 1/500.

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.
Target information above from: UniProt accession Q9UBU9 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 10482 Human
- Omim: 602647 Human
- SwissProt: Q9UBU9 Human
- Unigene: 523739 Human
- Unigene: 601546 Human
Alternative names
- DKFZp667O0311 antibody
- DmNXF1 antibody
- MEX67 antibody
- MEX67, yeast, homolog of antibody
- Mex67p antibody
- mRNA export factor TAP antibody
- Mvb1 antibody
- Nuclear RNA export factor 1 antibody
- Nuclear RNA export factor 1 homolog (S. cerevisiae) antibody
- nxf 1 antibody
- NXF1 antibody
- NXF1_HUMAN antibody
- Protein small bristles antibody
- Sbr antibody
- TAP antibody
- Tip associating protein antibody
- Tip-associated protein antibody
- Tip-associating protein antibody
see all

Western blot - Anti-NXF1 antibody [EPR8009] (ab129160)

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 - Anti-NXF1 antibody [EPR8009] (ab129160)

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.
Flow Cytometry - Anti-NXF1 antibody [EPR8009] (ab129160)

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).
Western blot - Anti-NXF1 antibody [EPR8009] (ab129160)

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
Immunocytochemistry/ Immunofluorescence - Anti-NXF1 antibody [EPR8009] (ab129160)

ab129160 at 1/250 dilution staining NXF1 in HepG2 cells by immunofluorescence.
Immunoprecipitation - Anti-NXF1 antibody [EPR8009] (ab129160)

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 lysate

For 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.
Immunoprecipitation - Anti-NXF1 antibody [EPR8009] (ab129160)

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 lysate

For 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.
Anti-NXF1 antibody [EPR8009] (ab129160)

Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

Publishing research using ab129160? Please let us know so that we can cite the reference in this datasheet.

ab129160 has been referenced in 3 publications.

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

Customer reviews and Q&As

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1-2 of 2 Abreviews or Q&A

Application

Immunoprecipitation

Sample

Human Cell lysate - whole cell (HeLa soluble lysate)

Total protein in input

5e+006 cells

Immuno-precipitation step

Protein A

Specification

HeLa soluble lysate

Abcam user community

Verified customer

Submitted Nov 30 2016

Application

Western blot

Sample

Human Cell lysate - whole cell (HeLa)

Gel Running Conditions

Reduced Denaturing (4-12% Bis-Tris)

Loading amount

100 µg

Treatment

siRNA verification

Specification

HeLa

Blocking step

Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Nov 29 2016

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Key features and details

Overview

Product name

Description

Host species

Tested Applications & Species

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Isotype control

Positive Controls

Recombinant Protein

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Sequence similarities

Domain

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

References (3)

Customer reviews and Q&As

Immunoprecipitation abreview for Anti-NXF1 antibody [EPR8009]

Western blot abreview for Anti-NXF1 antibody [EPR8009]