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遺伝子ノックアウト細胞株 検証済リコンビナントRabMAb

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

Reviews (2)Q&A (1)References (10)
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

製品の概要

  • 製品名

    Anti-Importin 9/RANBP9 antibody [EP1353Y]
    Importin 9/RANBP9 一次抗体 製品一覧

  • 製品の詳細

    Rabbit monoclonal [EP1353Y] to Importin 9/RANBP9

  • 由来種

    Rabbit

  • アプリケーション

    適用あり: WB, IHC-P, Flow Cyt (Intra)more details

  • 種交差性

    交差種: Human
    交差が予測される動物種: Mouse

  • 免疫原

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

  • ポジティブ･コントロール

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

  • 特記事項

    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.

製品の特性

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab52605の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
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.
特記事項
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.

ターゲット情報

  • 機能

    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.

  • 配列類似性

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

  • 細胞内局在

    Cytoplasm. Nucleus.
  • Information by UniProt

  • 参照データベース

    • 別名

      • 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

    画像

    • 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)

    プロトコール

    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

    参考文献 (10)

    ab52605 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab52605 は 10 報の論文で使用されています。

    • Kalita J  et al. On the asymmetric partitioning of nucleocytoplasmic transport - recent insights and open questions. J Cell Sci 134:N/A (2021). PubMed: 33912945
    • Palma M  et al. A role for AKT1 in nonsense-mediated mRNA decay. Nucleic Acids Res 49:11022-11037 (2021). PubMed: 34634811
    • 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

    レビューと Q&A

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    1-3 of 3 Abreviews or Q&A

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

     Good
    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

    投稿 Feb 19 2019

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

     Excellent
    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

    投稿 Feb 19 2019

    Question

    The data sheet for this RanBPM antibody shows a western blot indicating that this antibody recognizes a 115 Kda protein. RanBPM is 90 Kda. Could you explain the reason for this discrepancy? Also, could tell me what is the total amount and type of extract used to detect RanBPM with this antibody on this western blot.

    Read More
    Answer

    I can confirm that ab52605 is against RanBP9 not RanBPM, my apologies for the fact that RanBPM was added as an alternative name on the datasheet, this has been corrected immediately. The detected band's molecular weight matches the predicted MW detailed in the following database: http://www.expasy.org/uniprot/Q96P70 The protocol attached to the datasheet includes the extraction procedure, and I can confirm that 10 µg of fetal brain lysate was loaded (this tissue is purchased commercially and therefore we do not know the exact age I'm afraid). I hope this information will be useful.

    Read More

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