Recombinant Alexa Fluor® 647 Anti-HIF-1 alpha antibody [EPR16897] (ab208420)
![Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-HIF-1 alpha antibody [EPR16897] (ab208420)](https://www.abcam.com/ps/products/208/ab208420/Images/ab208420-268644-anti-hif-1-alpha-antibody-epr16897-alexa-fluor-647-immunofluorescence.jpg "Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-HIF-1 alpha antibody [EPR16897] (ab208420)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Alexa Fluor® 647 Rabbit monoclonal [EPR16897] to HIF-1 alpha
- Suitable for: ICC/IF
- Reacts with: Human
- Conjugation: Alexa Fluor® 647. Ex: 652nm, Em: 668nm
Related conjugates and formulations
Overview
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Product name
Alexa Fluor® 647 Anti-HIF-1 alpha antibody [EPR16897]
See all HIF-1 alpha primary antibodies -
Description
Alexa Fluor® 647 Rabbit monoclonal [EPR16897] to HIF-1 alpha -
Host species
Rabbit -
Conjugation
Alexa Fluor® 647. Ex: 652nm, Em: 668nm -
Tested applications
Suitable for: ICC/IFmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse
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Immunogen
Recombinant fragment within Human HIF-1 alpha aa 300-550. The exact immunogen sequence used to generate this antibody is proprietary information. If additional detail on the immunogen is needed to determine the suitability of the antibody for your needs, please contact our Scientific Support team to discuss your requirements.
Database link: Q16665 -
Positive control
- ICC/IF: DFO-treated HeLa cells
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General notes
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company. The Alexa Fluor® dye included in this product is provided under an intellectual property license from Life Technologies Corporation. As this product contains the Alexa Fluor® dye, the purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). As this product contains the Alexa Fluor® dye the sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are sold for use in research. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@thermofisher.com.
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: 0.02% Sodium azide
Constituents: PBS, 1% BSA, 30% Glycerol (glycerin, glycerine) -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR16897 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab208420 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 |
|---|---|---|
| ICC/IF |
1/50.
This product gave a positive signal in DFO-treated HeLa cells fixed with 4% formaldehyde (10 min) |
| Notes |
|---|
|
ICC/IF
1/50. This product gave a positive signal in DFO-treated HeLa cells fixed with 4% formaldehyde (10 min) |
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
- Entrez Gene: 15251 Mouse
- Omim: 603348 Human
- SwissProt: Q16665 Human
- SwissProt: Q61221 Mouse
- Unigene: 597216 Human
- Unigene: 3879 Mouse
- Unigene: 446610 Mouse
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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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Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-HIF-1 alpha antibody [EPR16897] (ab208420)
ab208420 staining HIF-1-alpha in HeLa cells +/- DFO (1nM, 24 hours). The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h.
The cells were then incubated overnight at +4°C with ab208420 at 1/50 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 488), at 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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![Alexa Fluor® 647 Anti-HIF-1 alpha antibody [EPR16897] (ab208420)](https://www.abcam.com/ps/products/208/ab208420/Images/ab208420-2-benefits-of-recombinant-antibodies.png)
Alexa Fluor® 647 Anti-HIF-1 alpha antibody [EPR16897] (ab208420)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (2)
ab208420 has been referenced in 2 publications.
- Luo X et al. Regulating Acidosis and Relieving Hypoxia by Platelet Membrane-Coated Nanoparticle for Enhancing Tumor Chemotherapy. Front Bioeng Biotechnol 10:885105 (2022). PubMed: 35646869
- Gilmore AC et al. An in vitro tumorigenesis model based on live-cell-generated oxygen and nutrient gradients. Commun Biol 4:477 (2021). PubMed: 33859337