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    importin-9ranbp9-antibody-ep1353y-ab52605.pdf

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Signal Transduction Protein Trafficking Nuclear Import / Export
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Importin 9/RANBP9 antibody [EP1353Y] (ab52605)

  • Datasheet
Reviews (2)Q&A (1)References (8)

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Western blot - Anti-Importin 9/RANBP9 antibody [EP1353Y] (ab52605)
  • Flow Cytometry (Intracellular) - Anti-Importin 9/RANBP9 antibody [EP1353Y] (ab52605)
  • Western blot - Anti-Importin 9/RANBP9 antibody [EP1353Y] (ab52605)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Importin 9/RANBP9 antibody [EP1353Y] (ab52605)
  • Anti-Importin 9/RANBP9 antibody [EP1353Y] (ab52605)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP1353Y] to Importin 9/RANBP9
  • Suitable for: WB, IHC-P, Flow Cyt (Intra)
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 647 Carrier Free

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Knockout
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Human IPO9 (Importin 9/RANBP9) knockout HeLa cell line (ab265352)

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Overview

  • Product name

    Anti-Importin 9/RANBP9 antibody [EP1353Y]
    See all Importin 9/RANBP9 primary antibodies
  • Description

    Rabbit monoclonal [EP1353Y] to Importin 9/RANBP9
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, Flow Cyt (Intra)more details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse
  • Immunogen

    Synthetic peptide within Human Importin 9/RANBP9 aa 1000 to the C-terminus (C terminal). The exact sequence is proprietary.
    Database link: Q96P70

  • Positive control

    • WB: HeLa and SK-BR-3 cell lysates; Fetal brain lysate. IHC-P: Human lymphoma tissue.
  • 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
    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.

    Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.

Properties

  • 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: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EP1353Y
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Protein Trafficking
    • Nuclear Import / Export
    • Cell Biology
    • Cell Cycle
    • Cell Division
    • Centromere
    • Epigenetics and Nuclear Signaling
    • Cell cycle
    • Chromosome Structure
    • Centromere
    • Epigenetics and Nuclear Signaling
    • Nuclear Signaling Pathways
    • Nuclear Receptors
    • Nuclear Pore Complex

Associated products

  • Alternative Versions

    • Alexa Fluor® 647 Anti-Importin 9/RANBP9 antibody [EP1353Y] (ab210416)
    • Anti-Importin 9/RANBP9 antibody [EP1353Y] - BSA and Azide free (ab247321)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab52605 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)
1/1000. Detects a band of approximately 115 kDa (predicted molecular weight: 115 kDa).
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt (Intra)
1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Notes
WB
1/1000. Detects a band of approximately 115 kDa (predicted molecular weight: 115 kDa).
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt (Intra)
1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Target

  • Function

    Functions in nuclear protein import as nuclear transport receptor. Serves as receptor for nuclear localization signals (NLS) in cargo substrates. Is thought to mediate docking of the importin/substrate complex to the nuclear pore complex (NPC) through binding to nucleoporin and the complex is subsequently translocated through the pore by an energy requiring, Ran-dependent mechanism. At the nucleoplasmic side of the NPC, Ran binds to the importin, the importin/substrate complex dissociates and importin is re-exported from the nucleus to the cytoplasm where GTP hydrolysis releases Ran. The directionality of nuclear import is thought to be conferred by an asymmetric distribution of the GTP- and GDP-bound forms of Ran between the cytoplasm and nucleus (By similarity). Mediates the nuclear import of H2B histone (By similarity), RPS7 and RPL18A. Prevents the cytoplasmic aggregation of RPS7 and RPL18A by shielding exposed basic domains. May also import H2A, H3, H4 histones (By similarity), RPL4 and RPL6.
  • Sequence similarities

    Belongs to the importin beta family.
    Contains 1 importin N-terminal domain.
  • Cellular localization

    Cytoplasm. Nucleus.
  • Target information above from: UniProt accession Q96P70 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 55705 Human
    • Entrez Gene: 226432 Mouse
    • SwissProt: Q96P70 Human
    • SwissProt: Q91YE6 Mouse
    • Unigene: 596014 Human
    • Unigene: 271642 Mouse
    • Alternative names

      • Imp9 antibody
      • Imp9a antibody
      • Imp9b antibody
      • Importin-9 antibody
      • Importin-9a antibody
      • Ipo9 antibody
      • IPO9_HUMAN antibody
      • Ran-binding protein 9 antibody
      • RanBP9 antibody
      see all

    Images

    • Western blot - Anti-Importin 9/RANBP9 antibody [EP1353Y] (ab52605)
      Western blot - Anti-Importin 9/RANBP9 antibody [EP1353Y] (ab52605)
      All lanes : Anti-Importin 9/RANBP9 antibody [EP1353Y] (ab52605) at 1/1000 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : IPO9 knockout HeLa cell lysate
      Lane 3 : SK-BR-3 cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 115 kDa



      Lanes 1-3: Merged signal (red and green). Green - ab52605. Red - loading control, ab8245 observed at 36 kDa. 

      ab52605 was shown to react with IPO9 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265352 (knockout cell lysate ab257483) was used. Wild-type and IPO9 knockout samples were subjected to SDS-PAGE. ab52605 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. 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 in 10000 dilution for 1 hour at room temperature before imaging.

    • Flow Cytometry (Intracellular) - Anti-Importin 9/RANBP9 antibody [EP1353Y] (ab52605)
      Flow Cytometry (Intracellular) - Anti-Importin 9/RANBP9 antibody [EP1353Y] (ab52605)

      Overlay histogram showing HeLa cells stained with ab52605 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab52605, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

       

    • Western blot - Anti-Importin 9/RANBP9 antibody [EP1353Y] (ab52605)
      Western blot - Anti-Importin 9/RANBP9 antibody [EP1353Y] (ab52605)
      Anti-Importin 9/RANBP9 antibody [EP1353Y] (ab52605) at 1/1000 dilution + Fetal brain lysate at 10 µg

      Secondary
      HRP-labelled goat anti-rabbit. at 1/2000 dilution

      Predicted band size: 115 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Importin 9/RANBP9 antibody [EP1353Y] (ab52605)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Importin 9/RANBP9 antibody [EP1353Y] (ab52605)

      Immunohistochemical staining of paraffin-embedded human lymphoma using ab52605 at a dilution of 1/50.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Anti-Importin 9/RANBP9 antibody [EP1353Y] (ab52605)
      Anti-Importin 9/RANBP9 antibody [EP1353Y] (ab52605)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    References (8)

    Publishing research using ab52605? Please let us know so that we can cite the reference in this datasheet.

    ab52605 has been referenced in 8 publications.

    • Trzaska C  et al. 2,6-Diaminopurine as a highly potent corrector of UGA nonsense mutations. Nat Commun 11:1509 (2020). PubMed: 32198346
    • Chatzifrangkeskou M  et al. RASSF1A is required for the maintenance of nuclear actin levels. EMBO J 38:e101168 (2019). PubMed: 31414556
    • Xue J  et al. Forkhead Box M1 Is Essential for Nuclear Localization of Glioma-associated Oncogene Homolog 1 in Glioblastoma Multiforme Cells by Promoting Importin-7 Expression. J Biol Chem 290:18662-70 (2015). PubMed: 26085085
    • Belin BJ  et al. DNA damage induces nuclear actin filament assembly by Formin -2 and Spire-½ that promotes efficient DNA repair. [corrected]. Elife 4:e07735 (2015). PubMed: 26287480
    • You P  et al. Expression and subcellular distribution of imp13 are regulated in brain development. In Vitro Cell Dev Biol Anim 49:346-53 (2013). IHC-P ; Mouse . PubMed: 23605716
    • Ratnakumar K  et al. ATRX-mediated chromatin association of histone variant macroH2A1 regulates a-globin expression. Genes Dev 26:433-8 (2012). WB ; Human . PubMed: 22391447
    • Lin W  et al. The roles of multiple importins for nuclear import of murine aristaless-related homeobox protein. J Biol Chem 284:20428-39 (2009). WB ; Human . PubMed: 19494118
    • Gast D  et al. The cytoplasmic part of L1-CAM controls growth and gene expression in human tumors that is reversed by therapeutic antibodies. Oncogene 27:1281-9 (2008). PubMed: 17952127

    Customer reviews and Q&As

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    1-2 of 2 Abreviews

    Western blot abreview for Anti-Importin 9/RANBP9 antibody [EP1353Y]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (USOS)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    20 µg
    Treatment
    siRNA
    Specification
    USOS
    Blocking step
    Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Feb 19 2019

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Importin 9 antibody [EP1353Y]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (U2OS)
    Permeabilization
    Yes - NP40
    Specification
    U2OS
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
    Fixative
    Formaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Feb 19 2019

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