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Products:Cardiovascular >> Hypoxia >> HIF
Overview
Product name
Anti-HIF1 alpha antibody - ChIP Grade
See all HIF1 alpha products (22) ...
Description
Rabbit polyclonal to HIF1 alpha - ChIP Grade
Tested applications
IHC-P, IHC-FoFr, WB, ICC/IF, ChIPmore details
Cross reactivity
Reacts with
Mouse, Rat, Chicken, Human, Monkey
Immunogen
Synthetic peptide, corresponding to amino acids 432-528 of Human HIF 1 alpha.
Properties
Form
Liquid
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage buffer
Preservative: 0.05% Sodium Azide
Constituents: Whole serum
Purity
Whole antiserum
Clonality
Polyclonal
Isotype
IgG
Research areas
Cancer >> Cancer Metabolism >> Response to hypoxia
Epigenetics and Nuclear Signaling >> ChIP'ing antibodies >> ChIP'ing antibodies
Cancer >> Invasion/microenvironment >> Hypoxia >> HIF
Epigenetics and Nuclear Signaling >> Cardiovascular/Immune >> Hypoxia >> HIF
Epigenetics and Nuclear Signaling >> Transcription >> Domain Families >> HLH / Leucine Zipper >> HLH
Cardiovascular >> Hypoxia >> HIF
Applications
Show applications key
Our Abpromise guarantee covers the use of ab2185 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
IHC-FoFr
WB
Application notes
ChIP: Use 25µl for 15 x 106 cells.
ICC/IF: 1/1000.
IHC-P: Use at an assay dependent dilution.
IHC-FoFr: Use at an assay dependent dilution.
WB: 1/500 - 1/1000. Predicted molecular weight: 120 kDa.
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
Target
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
modifications
In 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.
Target information above from: UniProt accessionQ16665
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Alternative names
- Basic helix loop helix PAS protein MOP1 antibody
- ARNT interacting protein antibody
- ARNT-interacting protein antibody
see all
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
see all
Anti-HIF1 alpha antibody - ChIP Grade images:
ChIP - HIF1 alpha antibody - ChIP Grade (ab2185)
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
Yolanda Cuevas, Hospital La Princesa.
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.
References for Anti-HIF1 alpha antibody - ChIP Grade (ab2185)
This product has been referenced in:
- Gaber Tet al. Macrophage migration inhibitory factor counterregulates dexamethasone-mediated suppression of hypoxia-inducible factor-1alpha function and differentially influences human CD4+ T cell proliferation under hypoxia. J Immunol 186:764-74 (2011).Read more (PubMed: 21169549) »
- Vanhorebeek Iet al. Insufficient activation of autophagy allows cellular damage to accumulate in critically ill patients. J Clin Endocrinol Metab 96:E633-45 (2011). WB; Human.Read more (PubMed: 21270330) »
See all 14 publications for this product
Publishing research using ab2185? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"