Anti-HIF1 alpha antibody - ChIP Grade (ab2185)

Overview

• Product nameAnti-HIF1 alpha antibody - ChIP GradeSee all HIF1 alpha primary antibodies ...
• Description
  Rabbit polyclonal to HIF1 alpha - ChIP Grade
• Tested applicationsIHC-P, IHC-FoFr, WB, ICC/IF, ChIP more details
• Species reactivity
  Reacts with: Mouse, Rat, Chicken, Human, Monkey
  
• Immunogen

  Synthetic peptide, corresponding to amino acids 432-528 of Human HIF 1 alpha.

Properties

• FormLiquid
• Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
• Storage bufferPreservative: 0.05% Sodium Azide
  Constituents: Whole serum
• PurityWhole antiserum
• Clonality Polyclonal
• IsotypeIgG
• Research Areas
  • Metabolism
  • Types of disease
  • Cancer

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

  • Metabolism
  • Pathways and Processes
  • Metabolism processes
  • Hypoxia

  • 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
  • ChIP'ing antibodies
  • ChIP'ing antibodies

  • 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: 374177 Chicken
  • Entrez Gene: 3091 Human
  • Entrez Gene: 15251 Mouse
  • Entrez Gene: 29560 Rat
  • Omim: 603348 Human
  • SwissProt: Q9YIB9 Chicken
  • SwissProt: Q16665 Human
  • SwissProt: Q61221 Mouse

  • SwissProt: O35800 Rat
  • Unigene: 597216 Human
  • Unigene: 3879 Mouse
  • Unigene: 446610 Mouse
  • Unigene: 10852 Rat

  see all

• 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

  see all

Anti-HIF1 alpha antibody - ChIP Grade images

• 

  ChIP - HIF1 alpha antibody - ChIP Grade (ab2185)Yolanda Cuevas, Hospital La Princesa.
  ChIP analysis of HIF1-alpha genomic sequences from HeLa cells cultivated in normoxic (N) or hypoxic (Hx) conditions, using a HIF1-alpha polyclonal antibody (ab2185). For a negative control, IgG was used and the input as a positive control in the subsequent PCR. Primers for known target genes were used HIF1 alpha, A. EPO and B. VEGF. Review by Yolanda Cuevas submitted 7 July 2004
  
• 

  Immunocytochemistry/ Immunofluorescence - HIF1 alpha antibody - ChIP Grade (ab2185)
  ICC/IF image of ab2185 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab2185, 1/1000 dilution) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
  
  

Protocols

• WB protocol using MEF cells- Hif1 alpha
• Nuclear Extract Preparation for HIF 1 alpha