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Products:Cardiovascular >> Hypoxia >> HIF

Anti-HIF1 alpha antibody [H1alpha67] - ChIP Grade (ab1)

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Overview

Product name

Anti-HIF1 alpha antibody [H1alpha67] - ChIP Grade
See all HIF1 alpha products (22) ...

Description

Mouse monoclonal [H1alpha67] to HIF1 alpha - ChIP Grade

Tested applications

IHC-P, IP, ChIP, IHC-Fr, ICC/IF, WBmore details

Cross reactivity

Reacts with

Human

Predicted to work with

Mouse, Rat, Sheep, Rabbit, Cow, Pig, Ferret, Monkey

Immunogen

Fusion protein: DVMLPSPN E KLQNINLAM SPLPTAETPK PLRSSADPA L NQEVALKL EP NPESLEL SFT MPQIQD QTPS PSDGS TRQSS PEPN SPSEYC FYV DSDMV, corresponding to amino acids 432-528 of Human HIF1 alpha

Positive control

This antibody gave a positive signal in a whole cell extract of DFO-treated HeLa cells. IHC-P on glioblastoma multiforme and also WB on nuclear extracts of a hypoxically or CoCl2 induced cell line such as MCF-7,PC12, 293, or MEF.

Properties

Form

Liquid

Storage instructions

Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.

Storage buffer

pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS

Concentration

Concentration information loading...

Purity

Protein G purified

Clonality

Monoclonal

Clone number

H1alpha67

Isotype

IgG2b

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

  • Western blot - Anti-HIF1 alpha antibody [H1alpha67] - ChIP Grade (ab1)Western blot - Anti-HIF1 alpha antibody [H1alpha67] - ChIP Grade (ab1) image (enlarge)

  • Western blot - Anti-HIF1 alpha antibody [H1alpha67] - ChIP Grade (ab1)Western blot - Anti-HIF1 alpha antibody [H1alpha67] - ChIP Grade (ab1) image (enlarge)

  • Immunocytochemistry/ Immunofluorescence - HIF1 alpha antibody [H1alpha67] (ab1)Immunocytochemistry/ Immunofluorescence - HIF1 alpha antibody [H1alpha67] (ab1) image (enlarge)

Applications

Show applications key

Our Abpromise guarantee covers the use of ab1 in the following tested applications.

The application notes include recommended starting dilutions; 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).

Information by UniProt

Alternative names

  • Basic helix loop helix PAS protein MOP1 antibody
  • ARNT interacting protein antibody
  • ARNT-interacting protein antibody
  • Basic-helix-loop-helix-PAS protein MOP1 antibody
  • bHLHe78 antibody
  • Class E basic helix-loop-helix protein 78 antibody
  • HIF 1A antibody
  • HIF 1alpha antibody
  • HIF-1-alpha antibody
  • HIF1 A antibody
  • HIF1 Alpha antibody
  • HIF1-alpha antibody
  • HIF1A antibody
  • HIF1A_HUMAN antibody
  • Hypoxia inducible factor 1 alpha antibody
  • Hypoxia inducible factor 1 alpha isoform I.3 antibody
  • Hypoxia inducible factor 1 alpha subunit antibody
  • Hypoxia inducible factor 1 alpha subunit basic helix loop helix transcription factor antibody
  • Hypoxia-inducible factor 1-alpha antibody
  • Member of PAS protein 1 antibody
  • Member of PAS superfamily 1 antibody
  • Member of the PAS Superfamily 1 antibody
  • MOP 1 antibody
  • MOP1 antibody
  • PAS domain-containing protein 8 antibody
  • PASD 8 antibody
  • PASD8 antibody
see all

Database links

see all

Anti-HIF1 alpha antibody [H1alpha67] - ChIP Grade images:

  Western blot - Anti-HIF1 alpha antibody [H1alpha67] - ChIP Grade (ab1)

Western blot - Anti-HIF1 alpha antibody [H1alpha67] - ChIP Grade (ab1)

All lanes : Anti-HIF1 alpha antibody [H1alpha67] - ChIP Grade (ab1) at 5 µg/ml

Lane 1 : Hela - Vehicle only (Negative Control) Whole Cell Lysate
Lane 2 : Hela - DFO treated (0.5mM, 24h) Whole Cell Lysate

Lysates/proteins at 25 µg per lane.

Secondary
Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed (ab97040) at 1/5000 dilution
developed using the ECL technique

Performed under reducing conditions.

Predicted band size : 92 kDa
Observed band size : 120 kDa (why is the actual band size different from the predicted?)
Additional bands at : 60 kDa. We are unsure as to the identity of these extra bands.

Exposure time : 20 minutes

  Western blot - Anti-HIF1 alpha antibody [H1alpha67] - ChIP Grade (ab1)

Western blot - Anti-HIF1 alpha antibody [H1alpha67] - ChIP Grade (ab1)

Anti-HIF1 alpha antibody [H1alpha67] - ChIP Grade (ab1) at 1/400 dilution + Human Cell lysate - whole cell (human lung adenocarcinoma cell line ADLC-5M2) treated for 16 hours with 100 micromolar deferoxamine (DFO) at 20 µg

Performed under reducing conditions.

Predicted band size : 92 kDa
Observed band size : 120 kDa (why is the actual band size different from the predicted?)


PVDF membrane was used and blocked for 16 hours in 5% milk.

This image is taken from an Abreview submitted by Mike Campa, no further information is known about this image

  Immunocytochemistry/ Immunofluorescence - HIF1 alpha antibody [H1alpha67] (ab1)

Immunocytochemistry/ Immunofluorescence - HIF1 alpha antibody [H1alpha67] (ab1)

ab1 at 1/200 dilution staining HIF1 alpha in human 293FT cells by Immunocytochemistry/ Immunofluorescence. Cells were fixed in formaldehyde, permeabilized in 0.5% Trition X-100 and blocked in 5% BSA for 1 hour at 25°C. The primary antibody was used at 1/200 dilution in PBS and incubated with sample at 4°C for 12 hours. An Alexa Fluor® 488 conjugated Goat polyclonal to mouse IgG was used as secondary at 1/500 dilution.

This Image is courtesy of an anonymous Abreview

See Abreview

  Immunohistochemistry (Frozen sections) - HIF1 alpha antibody [H1alpha67] (ab1)

Immunohistochemistry (Frozen sections) - HIF1 alpha antibody [H1alpha67] (ab1)

ab1 staining HIF1 alpha in mouse liver tissue section by Immunohistochemistry (Frozen sections). Tissue samples were fixed with formaldehyde and permeablized with 0.2% Triton-X100 before blocking with 2% BSA for 30 minutes at 20°C. The sample was incubated with primary antibody (1/200) for 9 hours at 4°C. An Alexa Fluor®555-conjugated Goat polyclonal to mouse IgG was used as secondary antibody at 1/200 dilution. DAPI was used to stain the cell nuclei (blue).

This image is courtesy of an anonymous Abreview

See Abreview

Protocols

  • WB protocol using MEF cells- Hif1 alpha
  • Nuclear Extract Preparation for HIF 1 alpha
  • Recommended IHC-P protocol with ab1

    • References for Anti-HIF1 alpha antibody [H1alpha67] - ChIP Grade (ab1)

    This product has been referenced in:

    • Ingraham CAet al. Matrix Metalloproteinase (MMP)-9 Induced by Wnt Signaling Increases the Proliferation and Migration of Embryonic Neural Stem Cells at Low O2 Levels. J Biol Chem 286:17649-57 (2011). WB; Rat.Read more (PubMed: 21460212) »
    • Bento CFet al. Methylglyoxal-induced imbalance in the ratio of vascular endothelial growth factor to angiopoietin 2 secreted by retinal pigment epithelial cells leads to endothelial dysfunction. Exp Physiol 95:955-70 (2010). WB, IHC-Fr; Human, Rat.Read more (PubMed: 20562294) »

    See all 33 publications for this product

    Publishing research using ab1? 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"