Anti-HIF1 alpha antibody [EP1215Y] (ab51608)

Overview

• Product nameAnti-HIF1 alpha antibody [EP1215Y]See all HIF1 alpha primary antibodies ...
• Description
  Rabbit monoclonal [EP1215Y] to HIF1 alpha
• Specificityab51608 recognizes HIF1 alpha.
• Tested applicationsICC/IF, ICC, Flow Cyt, WB, IHC-P more details
• Species reactivity
  Reacts with: Human
  
• Immunogen

  Synthetic peptide corresponding to residues near the C terminal of human HIF1 alpha.

• Positive controlRamos + CoCl2 lysate and liver carcinoma.
• General notesWe have mixed customer feedback towards the rat specificity so we are unable to confirm and guarantee its performance with rat samples. Please contact technical team for more information. Produced under U.S. Patent No. 5,675,063.

Properties

• FormLiquid
• Storage instructionsStore at -20°C. Stable for 12 months at -20°C
• Dissociation constant (Kd) Kd = 2.24 x 10^-10 M
• Storage bufferPBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%
• PurityTissue culture supernatant
• Clonality Monoclonal
• Clone numberEP1215Y
• IsotypeIgG
• Research Areas
  • Metabolism
  • Types of disease
  • Cancer

  • Metabolism
  • Pathways and Processes
  • Metabolism processes
  • Hypoxia

  • Metabolism
  • Pathways and Processes
  • Metabolic signaling pathways
  • Nucleotide metabolism

  • Cancer
  • Cancer Metabolism
  • Response to hypoxia

  • Cancer
  • Invasion/microenvironment
  • Hypoxia
  • HIF

  • Cardiovascular
  • Hypoxia
  • HIF

  • Epigenetics and Nuclear Signaling
  • Cardiovascular/Immune
  • Hypoxia
  • HIF

  • Epigenetics and Nuclear Signaling
  • Transcription
  • Domain Families
  • HLH / Leucine Zipper
  • HLH

Applications

Target

• FunctionFunctions 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 specificityExpressed 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 similaritiesContains 1 basic helix-loop-helix (bHLH) domain.
  Contains 1 PAC (PAS-associated C-terminal) domain.
  Contains 2 PAS (PER-ARNT-SIM) domains.
• DomainContains 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 localizationCytoplasm. Nucleus. Cytoplasmic in normoxia, nuclear translocation in response to hypoxia. Colocalizes with SUMO1 in the nucleus, under hypoxia.

Target information above from: UniProt accession Q16665 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
• Database links
  • Entrez Gene: 3091 Human
  • Omim: 603348 Human
  • SwissProt: Q16665 Human
  • Unigene: 597216 Human

• Alternative names
  ARNT interacting protein antibodyARNT-interacting protein antibodyBasic helix loop helix PAS protein MOP1 antibody

  Basic-helix-loop-helix-PAS protein MOP1 antibodybHLHe78 antibodyClass E basic helix-loop-helix protein 78 antibodyHIF 1A antibodyHIF 1alpha antibodyHIF-1-alpha antibodyHIF1 A antibodyHIF1 Alpha antibodyHIF1 antibodyHIF1-alpha antibodyHIF1A antibodyHIF1A_HUMAN antibodyHypoxia inducible factor 1 alpha antibodyHypoxia inducible factor 1 alpha isoform I.3 antibodyHypoxia inducible factor 1 alpha subunit antibodyHypoxia inducible factor 1 alpha subunit basic helix loop helix transcription factor antibodyHypoxia inducible factor 1, alpha subunit (basic helix loop helix transcription factor) antibodyHypoxia inducible factor1alpha antibodyHypoxia-inducible factor 1-alpha antibodyMember of PAS protein 1 antibodyMember of PAS superfamily 1 antibodyMember of the PAS Superfamily 1 antibodyMOP 1 antibodyMOP1 antibodyPAS domain-containing protein 8 antibodyPASD 8 antibodyPASD8 antibody

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Anti-HIF1 alpha antibody [EP1215Y] images

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  Western blot - HIF1 alpha antibody [EP1215Y] (ab51608)
  Anti-HIF1 alpha antibody [EP1215Y] (ab51608) at 1/2000 dilution + Ramos + CoCl2 lysate at 10 µg
  
  Secondary
  Goat anti-rabbit HRP at 1/2000 dilution
  
  Predicted band size : 93 kDa
  
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  Immunohistochemistry (Paraffin-embedded sections) - HIF1 alpha antibody [EP1215Y] (ab51608)
  Immunohistochemical staining of human liver carcinoma (paraffin section) using ab51608 at a dilution of 1/100.
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  Immunocytochemistry/ Immunofluorescence - Anti-HIF1 alpha antibody [EP1215Y] (ab51608)
  ab51608 staining HIF1α in HepG2 cells treated with baicalein (ab120723), by ICC/IF. Increase in HIF1α expression correlates with increased concentration of baicalein as described in literature.
  The cells were incubated at 37°C for 6h in media containing different concentrations of ab120723 (baicalein) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab51608 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody.
  
• 

  Flow Cytometry-Anti-HIF1 alpha antibody [EP1215Y](ab51608)
  Overlay histogram showing HeLa cells stained with ab51608 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab51608, 1/50 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.
  
  
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  Immunocytochemistry/ Immunofluorescence-Anti-HIF1 alpha antibody [EP1215Y](ab51608)
  Immunohistochemical analysis using ab51608 showing positive staining in Breast carcinoma tissue.
  
  
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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF1 alpha antibody [EP1215Y] (ab51608)
  Immunohistochemical analysis using ab51608 showing positive staining in Thyroid gland carcinoma tissue.
  
  
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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)-Anti-HIF1 alpha antibody [EP1215Y](ab51608)
  Immunohistochemical analysis using ab51608 showing positive staining in Colonic adenocarcinoma tissue.
  
  
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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)-Anti-HIF1 alpha antibody [EP1215Y](ab51608)
  Immunohistochemical analysis using ab51608 showing positive staining in Medulloblastoma tissue.
  
  
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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)-Anti-HIF1 alpha antibody [EP1215Y](ab51608)
  Immunohistochemical analysis using ab51608 showing positive staining in Glioma tissue.
  
  
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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)-Anti-HIF1 alpha antibody [EP1215Y](ab51608)
  Immunohistochemical analysis using ab51608 showing positive staining in Squamous cell cervical carcinoma tissue.
  
  
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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF1 alpha antibody [EP1215Y] (ab51608)
  Immunohistochemical analysis using ab51608 showing positive staining in Lung adenocarcinoma tissue.
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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF1 alpha antibody [EP1215Y] (ab51608)
  Immunohistochemical analysis using ab51608 showing positive staining in Normal tonsil tissue.
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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF1 alpha antibody [EP1215Y] (ab51608)
  Immunohistochemical analysis using ab51608 showing positive staining in Prostatic benign tumor tissue.
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  Other-Anti-HIF1 alpha antibody [EP1215Y](ab51608)
  Equilibrium disassociation constant (K_D)
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