Overview

  • Product name

  • Description

    Rabbit polyclonal to HIF-1 alpha
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, ELISA, WBmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide corresponding to Human HIF-1 alpha conjugated to keyhole limpet haemocyanin.
    Database link: Q16665

  • Positive control

    • Induced culture of E.coli (one shot Top10 competent cells). IHC-P: Human breast adenocarcinoma FFPE tissue sections
  • General notes

    HIF-1 alpha can be a difficult target to work with so we have compiled a summary of all the important information you need to know including useful tips. This can be found in the protocols tab or alternatively click here to download the English version and here to download the Mandarin.

    Under normoxic conditions HIF-1 alpha has a short half-life. It is largely undetectable in cells or tissues grown under normoxic conditions. It is stabilized only at O2 concentrations below 5% and upon stabilization under hypoxic conditions HIF-1 translocates to the nucleus. Therefore we recommend western blots using nuclear extracts and running Hypoxia treated samples as positive control (ab180880). Hypoxia can be induced with treatment using certain agents e.g. CoCl2 or DFO, etc. so proper sample preparation is critical.

Properties

Applications

Our Abpromise guarantee covers the use of ab82832 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-P 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ELISA Use at an assay dependent concentration.
WB 1/1000. Detects a band of approximately 107 kDa (predicted molecular weight: 93 kDa).

Target

  • Function

    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.
  • Tissue specificity

    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.
  • Sequence similarities

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

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

    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.
  • Cellular localization

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

  • Alternative names

    • 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

Images

  • All lanes : Anti-HIF-1 alpha antibody (ab82832) at 1/1000 dilution

    Lane 1 : uninduced culture of E.coli (one shot Top10 competent cells)
    Lane 2 : induced culture of E.coli (one shot Top10 competent cells)

    Secondary
    All lanes : anti-Rabbit IgG HRP at 1/1000 dilution

    Predicted band size: 93 kDa
    Additional bands at: 80 kDa. We are unsure as to the identity of these extra bands.



    Molecular weight of tag is 14kDa.
  • IHC image of HIF-1-alpha staining in Human breast adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica Bond system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab82832, 1/100 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

References

This product has been referenced in:

  • Liu W  et al. TNF-a inhibits xenograft tumor formation by A549 lung cancer cells in nude mice via the HIF-1a/VASP signaling pathway. Oncol Rep 41:2418-2430 (2019). Read more (PubMed: 30816548) »
  • Jin Q  et al. Hypoxia-inducible factor-1 signaling pathway influences the sensitivity of HCC827 cells to gefitinib. Oncol Lett 17:4034-4043 (2019). Read more (PubMed: 30881515) »
See all 23 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (femur head)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 0.01 M Citrate Buffer pH6.0
Permeabilization
Yes - 0.1%Triton X 100
Specification
femur head
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Apr 06 2016

Question
Answer

The supplying lab let me know that they do not have data for ab82832 regarding with HIF2a.

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Answer

Thank you for contacting Abcam.

The immunogen for ab104072 is the full length recombinant human HIF1 alpha protein, so it will be a mix of multiple species of antibody targeting different regions of the protein.

The immunogen for ab82832 are XXXXX, one of which is between residues XXXX.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

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Answer

Thank you for your enquiry. I would like to reassure you that our antibodies have all been tested successfully and would be covered by our guarantee in the applications listed on the datasheets. Therefore, any HIF1 alpha or HIF2 alpha antibody tested in the species and application you are using should be suitable for your applications and species. Also, although we produce many in-house antibodies, we obtain other antibodies from a wide range of sources. Therefore, comparison experiments will not often have been done. If you wish to be more certain, I can suggest choosing the antibody that has been best characterized. By this I mean the antibody that has been tested in the most applications and has the most data available (on the datasheet). I can confirm we have the following antibodies for HIF1 alpha tested in western blot, human and mouse. These would all be guaranteed for use in WB, hu and ms. ab1 Anti-HIF1 Mouse monoclonal alpha antibody [H1alpha67] - ChIP Grade Applications: ChIP, ICC/IF, IHC-Fr, IHC-P, IP, WB Reacts with: Cow, Ferret, Human, Monkey, Mouse, Pig, Rabbit, Rat, Sheep  (3)    Click here (or use the following: https://www.abcam.com/index.html?datasheet=1).   ab113642 Mouse monoclonal Anti-HIF1 alpha antibody [1A3] Applications: ELISA, ICC/IF, IHC-P, WB Reacts with: Human, Monkey, Mouse  (7)  Click here (or use the following: https://www.abcam.com/index.html?datasheet=113642).        ab65979  Rabbit polyclonal Anti-HIF1 alpha antibody - Carboxyterminal end Applications: IHC-P, WB Reacts with: Human, Mouse, Rat    Click here (or use the following: https://www.abcam.com/index.html?datasheet=65979).        ab82832 Rabbit polyclonal Anti-HIF1 alpha antibody Applications:ELISA, WB Reacts with: Human, Mouse  Click here (or use the following: https://www.abcam.com/index.html?datasheet=82832). We have the following antibodies tested and guaranteed in Hu and WB for HIF2 alpha: ab109731 Rabbit polyclonal Anti-HIF2 alpha antibody Applications: ICC, ICC/IF, IHC-P, WB Reacts with: Human, Zebrafish Predicted Mouse, Rat, Cow, Pig   Click here (or use the following: https://www.abcam.com/index.html?datasheet=109731).          ab20654 Rabbit polyclonal Anti-HIF2 alpha antibody Applications: ICC/IF, IHC-P, WB Reacts with: Human, Rat Predicted: Mouse Click here (or use the following: https://www.abcam.com/index.html?datasheet=20654). I am sorry to confirm that as far as we are aware, non of our HIF2 alpha antibodies have been tested with samples from mouse. All tested and guaranteed species cross-reactivity information is stated on our datasheets, and these are updated as soon as any new information is brought to our attention.  For the antibodies I have quoted above, the alignment of the immunogen with the mouse protein is known to be high. It is therefore likely that these antibodies will detect in mouse, although we cannot guarantee this without testing.  If you would like to test one of the HIF2 alpha antibodies in mouse, please contact me again by replying to this message prior to the purchase as you may be eligible for our testing discount program. Otherwise, we like to encourage all of our customers to submit an Abreview via the online product datasheet. We always appreciate customer feedback, whether positive or negative, and we make all product information available to researchers. Each Abreview earns Abpoints that can be used for discounts on future purchases or rewards such as Amazon.com gift certificates. To find out more about our Abreview system, please see the following link: https://www.abcam.com/abreviews I look forward to hearing from you. Should you have any further questions please do not hesitate to ask.    

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