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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

Anti-HIF-1 alpha 抗体 [54/HIF-1a] (ab279654)

Submit a review Submit a question References (3)
Western blot - Anti-HIF-1 alpha antibody [54/HIF-1a] (ab279654)
  • Western blot - Anti-HIF-1 alpha antibody [54] (ab279654)
  • Immunocytochemistry/ Immunofluorescence - Anti-HIF-1 alpha antibody [54/HIF-1a] (ab279654)
  • Anti-HIF-1 alpha antibody [54/HIF-1a] (ab279654)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Mouse monoclonal [54/HIF-1a] to HIF-1 alpha
  • Suitable for: ICC/IF, WB
  • 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-HIF-1 alpha antibody [54/HIF-1a]
    HIF-1 alpha 一次抗体 製品一覧

  • 製品の詳細

    Mouse monoclonal [54/HIF-1a] to HIF-1 alpha

  • 由来種

    Mouse

  • アプリケーション

    適用あり: ICC/IF, WBmore details
    適用なし: IHC-P

  • 種交差性

    交差種: Human

  • 免疫原

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • ポジティブ･コントロール

    • WB: HeLa treated with 1 mM DFO (Deferoxamine mesylate) for 24 hrs, whole cell lysate. HeLa treated with 0.5 mM CoCl2 for 6 hrs, whole cell lysate. ICC/IF: HeLa cells treated with 1 mM desferrioxamine for 24 hrs.

  • 特記事項

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

    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.

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ab279654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
ICC/IF
1/20.
WB
1/2000. Predicted molecular weight: 92 kDa.
特記事項
ICC/IF
1/20.
WB
1/2000. Predicted molecular weight: 92 kDa.
追加情報
Is unsuitable for IHC-P.

ターゲット情報

  • 機能

    Functions as a master transcriptional regulator of the adaptive response to hypoxia. Under hypoxic conditions activates the transcription of over 40 genes, including, erythropoietin, glucose transporters, glycolytic enzymes, vascular endothelial growth factor, and other genes whose protein products increase oxygen delivery or facilitate metabolic adaptation to hypoxia. Plays an essential role in embryonic vascularization, tumor angiogenesis and pathophysiology of ischemic disease. Binds to core DNA sequence 5'-[AG]CGTG-3' within the hypoxia response element (HRE) of target gene promoters. Activation requires recruitment of transcriptional coactivators such as CREBPB and EP300. Activity is enhanced by interaction with both, NCOA1 or NCOA2. Interaction with redox regulatory protein APEX seems to activate CTAD and potentiates activation by NCOA1 and CREBBP.

  • 組織特異性

    Expressed in most tissues with highest levels in kidney and heart. Overexpressed in the majority of common human cancers and their metastases, due to the presence of intratumoral hypoxia and as a result of mutations in genes encoding oncoproteins and tumor suppressors.

  • 配列類似性

    Contains 1 basic helix-loop-helix (bHLH) domain.
    Contains 1 PAC (PAS-associated C-terminal) domain.
    Contains 2 PAS (PER-ARNT-SIM) domains.

  • ドメイン

    Contains two independent C-terminal transactivation domains, NTAD and CTAD, which function synergistically. Their transcriptional activity is repressed by an intervening inhibitory domain (ID).

  • 翻訳後修飾

    In normoxia, is hydroxylated on Pro-402 and Pro-564 in the oxygen-dependent degradation domain (ODD) by EGLN1/PHD1 and EGLN2/PHD2. EGLN3/PHD3 has also been shown to hydroxylate Pro-564. The hydroxylated prolines promote interaction with VHL, initiating rapid ubiquitination and subsequent proteasomal degradation. Deubiquitinated by USP20. Under hypoxia, proline hydroxylation is impaired and ubiquitination is attenuated, resulting in stabilization.
    In normoxia, is hydroxylated on Asn-803 by HIF1AN, thus abrogating interaction with CREBBP and EP300 and preventing transcriptional activation. This hydroxylation is inhibited by the Cu/Zn-chelator, Clioquinol.
    S-nitrosylation of Cys-800 may be responsible for increased recruitment of p300 coactivator necessary for transcriptional activity of HIF-1 complex.
    Requires phosphorylation for DNA-binding.
    Sumoylated; by SUMO1 under hypoxia. Sumoylation is enhanced through interaction with RWDD3. Desumoylation by SENP1 leads to increased HIF1A stability and transriptional activity.
    Ubiquitinated; in normoxia, following hydroxylation and interaction with VHL. Lys-532 appears to be the principal site of ubiquitination. Clioquinol, the Cu/Zn-chelator, inhibits ubiquitination through preventing hydroxylation at Asn-803.
    The iron and 2-oxoglutarate dependent 3-hydroxylation of asparagine is (S) stereospecific within HIF CTAD domains.

  • 細胞内局在

    Cytoplasm. Nucleus. Cytoplasmic in normoxia, nuclear translocation in response to hypoxia. Colocalizes with SUMO1 in the nucleus, under hypoxia.
  • Information by UniProt

  • 参照データベース

    • 別名

      • ARNT interacting protein antibody
      • ARNT-interacting protein antibody
      • Basic helix loop helix PAS protein MOP1 antibody
      • Basic-helix-loop-helix-PAS protein MOP1 antibody
      • bHLHe78 antibody
      • Class E basic helix-loop-helix protein 78 antibody
      • HIF 1A antibody
      • HIF 1alpha antibody
      • HIF-1-alpha antibody
      • HIF-1alpha antibody
      • HIF-alpha antibody
      • HIF1 A antibody
      • HIF1 Alpha antibody
      • HIF1 antibody
      • HIF1-alpha antibody
      • HIF1A antibody
      • HIF1A_HUMAN antibody
      • hifla antibody
      • Hypoxia inducible factor 1 alpha antibody
      • Hypoxia inducible factor 1 alpha isoform I.3 antibody
      • Hypoxia inducible factor 1 alpha subunit antibody
      • Hypoxia inducible factor 1 alpha subunit basic helix loop helix transcription factor antibody
      • Hypoxia inducible factor 1, alpha subunit (basic helix loop helix transcription factor) antibody
      • Hypoxia inducible factor1alpha antibody
      • Hypoxia-inducible factor 1-alpha antibody
      • Member of PAS protein 1 antibody
      • Member of PAS superfamily 1 antibody
      • Member of the PAS Superfamily 1 antibody
      • MOP 1 antibody
      • MOP1 antibody
      • PAS domain-containing protein 8 antibody
      • PASD 8 antibody
      • PASD8 antibody
      see all

    画像

    • Western blot - Anti-HIF-1 alpha antibody [54/HIF-1a] (ab279654)
      Western blot - Anti-HIF-1 alpha antibody [54/HIF-1a] (ab279654)
      All lanes : Anti-HIF-1 alpha antibody [54/HIF-1a] (ab279654) at 1/2000 dilution

      Lane 1 : Wild-type HCT 116 Untreated DMOG Control cell lysate
      Lane 2 : Wild-type HCT 116 Treated DMOG (1 mM, 4 h) cell lysate
      Lane 3 : HIF1A knockout HCT 116 Untreated DMOG Control cell lysate
      Lane 4 : HIF1A knockout HCT 116 Treated DMOG (1 mM, 4 h) cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 92 kDa



      False colour image of Western blot: Anti-HIF-1 alpha antibody [54/HIF-1a] staining at 1/2000 dilution, shown in green; Rabbit anti-alpha Tubulin antibody [EP1332Y] (ab52866) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab279654 was shown to bind specifically to HIF-1 alpha. A band was observed at 110 kDa in treated wild-type HCT 116 cell lysates with no signal observed at this size in HIF1A knockout cell line ab255394 (knockout cell lysate ab263860). To generate this image, wild-type and HIF1A knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.

    • Western blot - Anti-HIF-1 alpha antibody [54] (ab279654)
      Western blot - Anti-HIF-1 alpha antibody [54] (ab279654)
      All lanes : Anti-HIF-1 alpha antibody [54/HIF-1a] (ab279654) at 1/2000 dilution

      Lane 1 : Untreated HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
      Lane 2 : HeLa treated with 1 mM DFO (Deferoxamine mesylate) for 24 hours whole cell lysate
      Lane 3 : Untreated HeLa whole cell lysate
      Lane 4 : HeLa treated with 0.5 mM CoCl2 for 6 hours whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/50000 dilution

      Predicted band size: 92 kDa
      Observed band size: 110 kDa why is the actual band size different from the predicted?



      Blocking and diluting buffer and concentration: 5% NFDM/TBST.

      The expression profile/molecular weight observed is consistent with what has been described in the literature (PMID: 9159130).

      Exposure time: 3 minutes.

    • Immunocytochemistry/ Immunofluorescence - Anti-HIF-1 alpha antibody [54/HIF-1a] (ab279654)
      Immunocytochemistry/ Immunofluorescence - Anti-HIF-1 alpha antibody [54/HIF-1a] (ab279654)

      Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling HIF-1 alpha with ab279654 at 1/20 dilution, followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green).

      Confocal image showing nuclear staining in HeLa cells treated with 1 mM desferrioxamine for 24 hrs.

      ab179513 anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) at 1/1000 dilution (Red). The nuclear counterstain was DAPI (Blue).

      Secondary antibody only controls: Secondary antibodies are ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution and ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) at 1/1000 dilution.

    • Anti-HIF-1 alpha antibody [54/HIF-1a] (ab279654)
      Anti-HIF-1 alpha antibody [54/HIF-1a] (ab279654)

    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 (3)

    ab279654 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。

    ab279654 は 3 報の論文で使用されています。

    • Jain L  et al. Differential Effects of Hypoxia versus Hyperoxia or Physoxia on Phenotype and Energy Metabolism in Human Chondrocytes from Osteoarthritic Compared to Macroscopically Normal Cartilage. Int J Mol Sci 24:N/A (2023). PubMed: 37108698
    • Meng F  et al. LncRNA LINC00525 activates HIF-1a through miR-338-3p / UBE2Q1 / ß-catenin axis to regulate the Warburg effect in colorectal cancer. Bioengineered 13:2554-2567 (2022). PubMed: 35156520
    • Zhang J  et al. Combining immune checkpoint blockade with ATP-based immunogenic cell death amplifier for cancer chemo-immunotherapy. Acta Pharm Sin B 12:3694-3709 (2022). PubMed: 36176905

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