Overview

  • Product name
    Anti-HIF1AN / FIH-1 antibody
    See all HIF1AN / FIH-1 primary antibodies
  • Description
    Rabbit polyclonal to HIF1AN / FIH-1
  • Host species
    Rabbit
  • Specificity
    The expected molecular weight of FIH is 40 kDa. A band corresponding to this size is seen on the western blot analysis, and is competed away by the addition of FIH blocking peptide. A smaller band is seen, which is also competed away by the blocking peptide. In the A431 extract a nonspecific band of approximately 50 kDa is seen, which is not diminished by the addition of blocking peptide.
  • Tested applications
    Suitable for: IHC-FoFr, ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Chicken, Human
  • Immunogen

    Synthetic peptide corresponding to Human HIF1AN/ FIH-1 aa 250 to the C-terminus (internal sequence) conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab12306)

  • Positive control
    • Recombinant Human HIF1AN / FIH-1 protein (ab86916) can be used as a positive control in WB. This antibody gave a positive signal in the following whole cell lysates: HeLa; Jurkat; A431; Hek293 and Hela Nuclear lysate.
  • General notes

    Previously labelled as HIF1AN.

Properties

Applications

Our Abpromise guarantee covers the use of ab12289 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
IHC-FoFr
ICC/IF
WB
  • Application notes
    ICC/IF: Use at a concentration of 5 µg/ml.
    WB: 1/500. Detects a band of approximately 40 kDa (predicted molecular weight: 40 kDa). Can be blocked with FIH peptide (ab12306).


    Not yet tested in other applications.
    Optimal dilutions/concentrations should be determined by the end user.
  • Target

    • Function
      Hydroxylates HIF-1 alpha at 'Asp-803' in the C-terminal transactivation domain (CAD). Functions as an oxygen sensor and, under normoxic conditions, the hydroxylation prevents interaction of HIF-1 with transcriptional coactivators including Cbp/p300-interacting transactivator. Involved in transcriptional repression through interaction with HIF1A, VHL and histone deacetylases.
    • Sequence similarities
      Contains 1 JmjC domain.
    • Cellular localization
      Nucleus.
    • Information by UniProt
    • Database links
    • Alternative names
      • DKFZp762F1811 antibody
      • Factor inhibiting HIF-1 antibody
      • Factor inhibiting HIF1 antibody
      • FIH 1 antibody
      • FIH-1 antibody
      • FIH1 antibody
      • FLJ20615 antibody
      • FLJ22027 antibody
      • HIF1AN antibody
      • HIF1N_HUMAN antibody
      • Hypoxia inducible factor 1 alpha inhibitor antibody
      • Hypoxia inducible factor 1 alpha subunit inhibitor antibody
      • Hypoxia inducible factor asparagine hydroxylase antibody
      • Hypoxia-inducible factor 1-alpha inhibitor antibody
      • Hypoxia-inducible factor asparagine hydroxylase antibody
      • Peptide aspartate beta dioxygenase antibody
      see all

    Images

    • All lanes : Anti-HIF1AN / FIH-1 antibody (ab12289) at 1/500 dilution

      Lane 1 : Hela nuclear extract
      Lanes 2 & 7 : Hela whole cell lysate
      Lane 3 : A431 whole cell lysate
      Lane 4 : Jurkat whole cell lysate
      Lane 5 : Hek293 cell lysate
      Lane 6 : Hela nuclear extract with FIH blocking peptide at 1 µg/ml
      Lane 8 : A431 whole cell lysate with FIH blocking peptide at 1 µg/ml
      Lane 9 : Jurkat whole cell lysate with FIH blocking peptide at 1 µg/ml
      Lane 10 : Hek293 whole cell lysate with FIH blocking peptide at 1 µg/ml

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat polyclonal to rabbit IgG H&L (HRP) at 1/5000 dilution

      Predicted band size: 40 kDa
      Observed band size: 40 kDa


      Exposure time: 5 minutes


      Lane 1: 20 µg of HeLa nuclear extract.
      Lane 2: 20 µg of HeLa whole cell lysate.
      Lane 3: 20 µg of A431 whole cell lysate.
      Lane 4: 20 µ

    • ICC/IF image of ab12289 stained human Hek293 cells. The cells were 4% PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab12289, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).

    References

    This product has been referenced in:
    • Sallais J  et al. Factor inhibiting HIF1-A novel target of SUMOylation in the human placenta. Oncotarget 8:114002-114018 (2017). Read more (PubMed: 29371964) »
    • Yang Y  et al. A novel fumarate hydratase-deficient HLRCC kidney cancer cell line, UOK268: a model of the Warburg effect in cancer. Cancer Genet 205:377-90 (2012). Read more (PubMed: 22867999) »
    See all 4 Publications for this product

    Customer reviews and Q&As

    1-3 of 3 Abreviews or Q&A

    Answer

    Thanks for your call yesterday and for your patience. We have two HIF1AN antibodies that have been tested with mouse samples and are guaranteed to work in mouse: ab92304 and ab63163- https://www.abcam.com/HIF1AN-antibody-ab63163.html https://www.abcam.com/HIF1AN-antibody-EPR3659-ab92304.html I did not find any data showing these antibodies used against knockout samples, but I would not expect them to cross-react with other proteins. Regarding ab12289, do you have any testing data such as a Western blot image that you could send? What kind of mouse tissue was used in this assay? I look forward to hearing from you. Please let me know if you have any questions or if there is anything else that we can do for you.

    Read More
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HEK293, Jurkat, Raw264)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    20 µg
    Specification
    HEK293, Jurkat, Raw264
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

    Mr. Antibody Solutions

    Verified customer

    Submitted Feb 24 2009

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa and SK-N-SH)
    Specification
    HeLa and SK-N-SH
    Fixative
    Paraformaldehyde

    Dr. Kirk Mcmanus

    Verified customer

    Submitted Jul 24 2007

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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