HRP Anti-HIF-1 alpha antibody [ESEE122] (ab215773)
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - HRP Anti-HIF-1 alpha antibody [ESEE122] (ab215773)](https://www.abcam.com/ps/products/215/ab215773/Images/ab215773-291653-anti-hif-1-alpha-antibody-esee122-hrp-immunohistochemistry.jpg "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - HRP Anti-HIF-1 alpha antibody [ESEE122] (ab215773)")
Key features and details
- HRP Mouse monoclonal [ESEE122] to HIF-1 alpha
- Suitable for: IHC-P, WB
- Reacts with: Human
- Conjugation: HRP
- Isotype: IgG1
Overview
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Product name
HRP Anti-HIF-1 alpha antibody [ESEE122]
See all HIF-1 alpha primary antibodies -
Description
HRP Mouse monoclonal [ESEE122] to HIF-1 alpha -
Host species
Mouse -
Conjugation
HRP -
Tested applications
Suitable for: IHC-P, WBmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment within Human HIF-1 alpha aa 300-550. The exact sequence is proprietary.
Database link: Q16665 -
Positive control
- HeLa nuclear extract IHC-P: normal human colon tissue sections
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General notes
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. Store In the Dark. -
Storage buffer
pH: 7.40
Preservative: 0.1% 10% Proclin 300 Solution
Constituents: 30% Glycerol, 1% BSA, PBS -
Concentration information loading... -
Purity
Immunogen affinity purified -
Clonality
Monoclonal -
Clone number
ESEE122 -
Myeloma
NS1 -
Isotype
IgG1 -
Research areas
Associated products
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Alternative Versions
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Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab215773 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 Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. | |
| WB | 1/5000. Detects a band of approximately 92 kDa (predicted molecular weight: 92 kDa). Ensure cell lysis occurs quickly if removed from hypoxia. Ultrasonic lysis is recommended to enrich more nuclear lysate. Positive Control: Hypoxic samples such as HeLa-DFO treated whole cell lysate ab116322. For a stronger signal, HeLa-DFO treated nuclear extracts are recommended ab180880. The cell fractionation kit can also be purchased separately ab109719. Negative Control: Most normal tissues or cells, other than kidney or heart. The observed band size of HIF-1 alpha is not exactly as predicted due to the different forms of HIF-1 alpha below: |
Target
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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
modificationsIn 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
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Database links
- Entrez Gene: 3091 Human
- Omim: 603348 Human
- SwissProt: Q16665 Human
- Unigene: 597216 Human
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Alternative names
- ARNT interacting protein antibody
- ARNT-interacting protein antibody
- Basic helix loop helix PAS protein MOP1 antibody
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - HRP Anti-HIF-1 alpha antibody [ESEE122] (ab215773)
IHC image of HIF-1-alpha staining in a section of formalin-fixed paraffin-embedded normal human colon*, performed on a Leica BOND™. The section was pre-treated using heat mediated antigen retrieval with Tris/EDTA buffer (pH9, epitope retrieval solution 2) for 20mins. The section was then incubated with ab215773, 1/100 dilution, for 15 mins at room temperature. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.
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.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
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![Western blot - HRP Anti-HIF-1 alpha antibody [ESEE122] (ab215773)](https://www.abcam.com/ps/products/215/ab215773/Images/ab215773-283999-anti-hif-1-alpha-antibody-esee122-hrp-western-blot.jpg)
Western blot - HRP Anti-HIF-1 alpha antibody [ESEE122] (ab215773)All lanes : HRP Anti-HIF-1 alpha antibody [ESEE122] (ab215773) at 1/5000 dilution
Lane 1 : HeLa nuclear extract lysate
Lane 2 : Hela-DFO treated (0.5mM, 24h) Nuclear Lysate
Lysates/proteins at 10 µg per lane.
Performed under reducing conditions.
Predicted band size: 92 kDa
Observed band size: 92 kDa
Exposure time: 2 minutesThis blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab215773 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.
Protocols
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.
References (0)
ab215773 has not yet been referenced specifically in any publications.
