Anti-HIF-1 alpha antibody (ab216842)
Key features and details
- Rabbit polyclonal to HIF-1 alpha
- Suitable for: WB, ICC/IF, Flow Cyt (Intra)
- Reacts with: Mouse, Human
- Isotype: IgG
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Overview
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Product name
Anti-HIF-1 alpha antibody
See all HIF-1 alpha primary antibodies -
Description
Rabbit polyclonal to HIF-1 alpha -
Host species
Rabbit -
Tested applications
Suitable for: WB, ICC/IF, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
Synthetic peptide within Human HIF-1 alpha aa 350-450 conjugated to keyhole limpet haemocyanin. 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
- WB: Hela, A431, U251, HepG2, HL60 and U-87MG cell lysate; ICC/IF: Hela cells. Flow Cyt (Intra): Mouse spleen cells.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, 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 long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Proclin 300
Constituents: 50% Glycerol (glycerin, glycerine), 1% BSA, 48.98% TBS, 1X -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Assay kits
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab216842 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 |
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WB |
1/300 - 1/1000. 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: |
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ICC/IF |
1/50 - 1/200.
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Flow Cyt (Intra) |
1/20 - 1/100.
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Notes |
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WB
1/300 - 1/1000. 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: |
ICC/IF
1/50 - 1/200. |
Flow Cyt (Intra)
1/20 - 1/100. |
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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All lanes : Anti-HIF-1 alpha antibody (ab216842) at 1/1000 dilution
Lane 1 : Hela cell lysate
Lane 2 : A431 cell lysate
Lane 3 : U251 cell lysate
Lane 4 : HepG2 cell lysate
Secondary
All lanes : Conjugated secondary antibody at 1/20000 dilution
Predicted band size: 92 kDaCell lysates were first probed with primary antibody (ab216842) at 4°C overnight then incubated with conjugated secondary antibody for 60 min at 37°C.
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Hela cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with HIF-1 Alpha polyclonal Antibody (1/100, ab216842), 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.
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Mouse spleen cells were fixed with 4% PFA for 10min at room temperature, permeabilized with 90% ice-cold methanol for 20 min at -20℃, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with HIF-1 Alpha Antibody (ab216842) at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).
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Anti-HIF-1 alpha antibody (ab216842) at 1/300 dilution + HL60 cell lysate
Secondary
Goat Anti-Rabbit IgG Antibody (H+L), HRP at 1/5000 dilution
Predicted band size: 92 kDaCell lysate was first probed with primary antibody (ab216842) at 4°C overnight then incubated with conjugated secondary antibody for 90 min at 37°C.
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Anti-HIF-1 alpha antibody (ab216842) at 1/1000 dilution + U-87MG cell lysate
Secondary
Conjugated secondary antibody at 1/20000 dilution
Predicted band size: 92 kDaCell lysate was first probed with primary antibody (ab216842) at 4°C overnight then incubated with conjugated secondary antibody for 60 min at 37°C.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (38)
ab216842 has been referenced in 38 publications.
- Kim BH et al. Restoration of Homeostasis in the Tracheal Mucosa After Thyroid Surgery in a Rat Model. In Vivo 36:161-169 (2022). PubMed: 34972711
- Ni H et al. Cardamonin attenuates cerebral ischemia/reperfusion injury by activating the HIF-1α/VEGFA pathway. Phytother Res 36:1736-1747 (2022). PubMed: 35142404
- Liu Y et al. RIG-I Promotes Cell Viability, Colony Formation, and Glucose Metabolism and Inhibits Cell Apoptosis in Colorectal Cancer by NF-κB Signaling Pathway. Dis Markers 2022:1247007 (2022). PubMed: 35242239
- Liu Y et al. Hypoxia During the Consolidation Phase of Distraction Osteogenesis Promotes Bone Regeneration. Front Physiol 13:804469 (2022). PubMed: 35283791
- Du P et al. Mucin-fused myeloid-derived growth factor (MYDGF164) exhibits a prolonged serum half-life and alleviates fibrosis in chronic kidney disease. Br J Pharmacol 179:4136-4156 (2022). PubMed: 35393682