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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-eIF4EBP1 抗体 [Y330] (ab32130)

Submit a review Submit a question References (9)
Western blot - Anti-eIF4EBP1 antibody [Y330] (ab32130)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF4EBP1 antibody [Y330] (ab32130)
  • Flow Cytometry (Intracellular) - Anti-eIF4EBP1 antibody [Y330] (ab32130)
  • Western blot - Anti-eIF4EBP1 antibody [Y330] (ab32130)
  • Western blot - Anti-eIF4EBP1 antibody [Y330] (ab32130)
  • Anti-eIF4EBP1 antibody [Y330] (ab32130)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [Y330] to eIF4EBP1
  • Suitable for: WB, IHC-P, Flow Cyt (Intra)
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

製品の概要

  • 製品名

    Anti-eIF4EBP1 antibody [Y330]
    eIF4EBP1 一次抗体 製品一覧

  • 製品の詳細

    Rabbit monoclonal [Y330] to eIF4EBP1

  • 由来種

    Rabbit

  • 特異性

    ab32130 recognises 4E-BP1 (eIF4E-binding protein) also known as PHAS.

  • アプリケーション

    適用あり: WB, IHC-P, Flow Cyt (Intra)more details
    適用なし: ICC/IF

  • 種交差性

    交差種: Human

  • 免疫原

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • ポジティブ･コントロール

    • WB: HeLa, HEK-293 cell lysate, HAP1 and K562 cell lysate, IHC-P: Human prostate carcinoma, Flow Cyt (intra): HeLa cells

  • 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.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these 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ab32130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
1/1000. Detects a band of approximately 15-20 kDa (predicted molecular weight: 12 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.
特記事項
WB
1/1000. Detects a band of approximately 15-20 kDa (predicted molecular weight: 12 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.
追加情報
Is unsuitable for ICC/IF.

ターゲット情報

  • 機能

    Regulates eIF4E activity by preventing its assembly into the eIF4F complex. Mediates the regulation of protein translation by hormones, growth factors and other stimuli that signal through the MAP kinase and mTORC1 pathways.

  • 配列類似性

    Belongs to the eIF4E-binding protein family.

  • 翻訳後修飾

    Phosphorylated on serine and threonine residues in response to insulin, EGF and PDGF. Phosphorylation at Thr-37, Thr-46, Ser-65 and Thr-70 is regulated by mTORC1. Phosphorylated upon DNA damage, probably by ATM or ATR.
  • Information by UniProt

  • 参照データベース

    • 別名

      • 4E-BP1 antibody
      • 4EBP1 antibody
      • 4EBP1_HUMAN antibody
      • BP 1 antibody
      • eIF4E binding protein 1 antibody
      • eIF4E-binding protein 1 antibody
      • Eif4ebp1 antibody
      • Eukaryotic translation initiation factor 4E-binding protein 1 antibody
      • PHAS-I antibody
      • PHASI antibody
      • Phosphorylated heat- and acid-stable protein regulated by insulin 1 antibody
      see all

    画像

    • Western blot - Anti-eIF4EBP1 antibody [Y330] (ab32130)
      Western blot - Anti-eIF4EBP1 antibody [Y330] (ab32130)
      All lanes : Anti-eIF4EBP1 antibody [Y330] (ab32130) at 1/1000 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : EIF4EBP1 knockout HeLa cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 12 kDa
      Observed band size: 13 kDa why is the actual band size different from the predicted?



      Lanes 1-2: Merged signal (red and green). Green - ab32130 observed at 13 kDa. Red - loading control ab8245 observed at 37 kDa.

       ab32130 Anti-eIF4EBP1 antibody [Y330] was shown to specifically react with eIF4EBP1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264784 (knockout cell lysate ab257146) was used. Wild-type and eIF4EBP1 knockout samples were subjected to SDS-PAGE. ab32130 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 20000 dilution for 1 hour at room temperature before imaging.

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF4EBP1 antibody [Y330] (ab32130)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF4EBP1 antibody [Y330] (ab32130)

      ab32130, staining human prostate carcinoma by immunohistochemistry, Paraffin embedded tissue

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

    • Flow Cytometry (Intracellular) - Anti-eIF4EBP1 antibody [Y330] (ab32130)
      Flow Cytometry (Intracellular) - Anti-eIF4EBP1 antibody [Y330] (ab32130)

      Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling eIF4EBP1 with purified ab32130 at 1/1000 dilution (1 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 (Left).
      Low expression in mitotic phase was consistent with literature. (PMID: 11553333)

    • Western blot - Anti-eIF4EBP1 antibody [Y330] (ab32130)
      Western blot - Anti-eIF4EBP1 antibody [Y330] (ab32130)
      All lanes : Anti-eIF4EBP1 antibody [Y330] (ab32130) at 1/1000 dilution

      Lane 1 : Wild-type HAP1 cell lysate
      Lane 2 : eIF4EBP1 knockout HAP1 cell lysate
      Lane 3 : Hek293 cell lysate
      Lane 4 : K562 cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 12 kDa



      Lanes 1 - 4: Merged signal (red and green). Green - ab32130 observed at 20 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab32130 was shown to specifically react with eIF4EBP1 when eIF4EBP1 knockout samples were used. Wild-type and eIF4EBP1 knockout samples were subjected to SDS-PAGE. ab32130 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/2000 respectively and incubated overnight at 4°C. 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 1/10000 dilution for 1 h at room temperature before imaging.

    • Western blot - Anti-eIF4EBP1 antibody [Y330] (ab32130)
      Western blot - Anti-eIF4EBP1 antibody [Y330] (ab32130)
      Anti-eIF4EBP1 antibody [Y330] (ab32130) at 1/1000 dilution + K562 cell lysate

      Predicted band size: 12 kDa
      Observed band size: 18 kDa why is the actual band size different from the predicted?

    • Anti-eIF4EBP1 antibody [Y330] (ab32130)
      Anti-eIF4EBP1 antibody [Y330] (ab32130)

    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 (9)

    ab32130 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。

    ab32130 は 9 報の論文で使用されています。

    • Liu L  et al. Se improves GPX4 expression and SOD activity to alleviate heat-stress-induced ferroptosis-like death in goat mammary epithelial cells. Anim Cells Syst (Seoul) 25:283-295 (2021). PubMed: 34745435
    • Guo Y  et al. Spontaneous formation of neutrophil extracellular traps is associated with autophagy. Sci Rep 11:24005 (2021). PubMed: 34907287
    • Batool A  et al. Eukaryotic Initiation Factor 4E phosphorylation acts a switch for its binding to 4E-BP1 and mRNA cap assembly. Biochem Biophys Res Commun 527:489-495 (2020). PubMed: 32336547
    • Hori Y  et al. Analysis of mTOR pathway expression in lymphatic malformation and related diseases. Pathol Int 70:323-329 (2020). PubMed: 32067331
    • Wu X  et al. PI3K/AKT/mTOR pathway inhibitors inhibit the growth of melanoma cells with mTOR H2189Y mutations in vitro. Cancer Biol Ther 19:584-589 (2018). PubMed: 29708815
    • Wang W  et al. Rasfonin promotes autophagy and apoptosis via upregulation of reactive oxygen species (ROS)/JNK pathway. Mycology 7:64-73 (2016). PubMed: 30123617
    • Lu Q  et al. Akt inhibition attenuates rasfonin-induced autophagy and apoptosis through the glycolytic pathway in renal cancer cells. Cell Death Dis 6:e2005 (2015). Human . PubMed: 26633711
    • Hu Y  et al. MiR-99a antitumor activity in human breast cancer cells through targeting of mTOR expression. PLoS One 9:e92099 (2014). WB ; Human . PubMed: 24637915
    • Fukuda A  et al. Growth-dependent podocyte failure causes glomerulosclerosis. J Am Soc Nephrol 23:1351-63 (2012). PubMed: 22773827

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