Recombinant
RabMAb

Recombinant Anti-NXF1 antibody [EPR8009] (ab129160)

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Overview

  • Product name

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

  • Description

    Rabbit monoclonal [EPR8009] to NXF1

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IP, Flow Cyt, ICC/IFmore details

  • Species reactivity

    Reacts with: Human

  • 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. FC: 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.

Properties

Applications

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
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.

Target

  • 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

  • Database links

    • 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

    Images

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

    References

    This product has been referenced in:

    See 1 Publication for this product

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

    Customer reviews and Q&As

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    1-2 of 2 Abreviews or Q&A

    Immunoprecipitation abreview for Anti-NXF1 antibody [EPR8009]

     Excellent
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Nov 30 2016

    Western blot abreview for Anti-NXF1 antibody [EPR8009]

     Excellent
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Nov 29 2016

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