Overview

  • Product name
    Anti-HIF-1 alpha antibody [1A3]
    See all HIF-1 alpha primary antibodies
  • Description
    Mouse monoclonal [1A3] to HIF-1 alpha
  • Tested applications
    Suitable for: Flow Cyt, WB, ELISA, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human, Monkey
  • Immunogen

    Recombinant fragment corresponding to Human HIF-1-alpha (C terminal).

  • Positive control
    • Cos7, Hela, Jurkat, RAJI and NIH/3T3 cell lysate. HeLa cells. Liver, lung, stomach and brain cancer tissue.
  • General notes

    Under normoxic conditions HIF-1 alpha has a short half-life. It is largely undetectable in cells or tissues grown under normoxic conditions. It is stabilized only at O2 concentrations below 5% and upon stabilization under hypoxic conditions HIF-1 translocates to the nucleus. Therefore we recommend western blots using nuclear extracts and running Hypoxia treated samples as positive control (ab180880). Hypoxia can be induced with treatment using certain agents e.g. CoCl2 or DFO, etc. so proper sample preparation is critical.

    This product was changed from ascites to supernatant. Lot no’s high than GR206357-17 are from Tissue Culture Supernatant

Properties

Applications

Our Abpromise guarantee covers the use of ab113642 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
Flow Cyt 1/50 - 1/100.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

WB 1/500 - 1/2000. Predicted molecular weight: 93 kDa.
ELISA 1/10000.
IHC-P 1/200 - 1/1000.
ICC/IF 1/200 - 1/1000.

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.
  • Information by UniProt
  • Database links
  • Alternative names
    • ARNT interacting protein antibody
    • ARNT-interacting protein antibody
    • Basic helix loop helix PAS protein MOP1 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 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 subunit (basic helix loop helix transcription factor) antibody
    • Hypoxia inducible factor1alpha 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

Anti-HIF-1 alpha antibody [1A3] images

  • Immunocytochemistry/Immunofluorescence analysis of HeLa cells labeling HIF1 alpha (green) using ab113642 at 1/200. Blue: DRAQ5 fluorescent DNA dye. Red: Actin filaments labeled with Alexa Fluor® 555 phalloidin.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human liver cancer tissue sections labeling HIF-1-alpha with ab113642 at 1/200 with DAB staining.

  • All lanes : Anti-HIF-1 alpha antibody [1A3] (ab113642) at 1/500 dilution

    Lane 1 : Cos7 cell lysate
    Lane 2 : HeLa cell lysate
    Lane 3 : Jurkat cell lysate
    Lane 4 : Raji cell lysate
    Lane 5 : NIH 3T3 cell lysate


    Predicted band size : 93 kDa
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human lung cancer tissue sections labeling HIF-1-alpha with ab113642 at 1/200 with DAB staining.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human stomach cancer tissue sections labeling HIF-1-alpha using ab113642 at 1/200 with DAB staining.

  • Immunohistochemistry (Formalin-PFA-fixed paraffin-embedded sections) analysis Human brain tumor tissue sections labeling HIF-1-alpha with ab113642 at 1/200 with DAB staining.

  • Overlay histogram showing HeLa cells stained with ab113642 (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 (ab113642, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • Elisa using ab113642

References for Anti-HIF-1 alpha antibody [1A3] (ab113642)

This product has been referenced in:
  • Shao C  et al. Expression and significance of GRHL2 in esophageal cancer. Onco Targets Ther 10:2025-2031 (2017). IHC-P ; Human . Read more (PubMed: 28435293) »
  • Han J  et al. Interleukin-6 promotes tumor progression in colitis-associated colorectal cancer through HIF-1a regulation. Oncol Lett 12:4665-4670 (2016). IHC-P ; Human . Read more (PubMed: 28105173) »

See all 13 Publications for this product

Product Wall

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HEK293)
Permeabilization
Yes - 0.1% Triton X100
Specification
HEK293
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Jul 31 2017

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (4-15%)
Sample
Mouse Cell lysate - nuclear (infected with toxoplasma for 18hrs)
Specification
infected with toxoplasma for 18hrs
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Feb 05 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (B16F10 melanoma cells)
Loading amount
30 µg
Specification
B16F10 melanoma cells
Gel Running Conditions
Non-reduced Denaturing (8%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Username

Ms. Seontae Kim

Verified customer

Submitted Jan 02 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (B16F10 melanoma)
Specification
B16F10 melanoma
Fixative
4% Formalin
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: autoclaved for 10min
Permeabilization
Yes - 0.1% SDS
Blocking step
BSA as blocking agent for 15 minute(s) · Concentration: 1% · Temperature: 25°C
Username

Ms. Seontae Kim

Verified customer

Submitted Dec 19 2012

Thank you for contacting us.

Unfortunately, because we carry over 90,000 products, it isn't feasible for us to keep small sample sizes of our products, and therefore I am not able to offer a test sample of any of the antibodies requested. Read More

Thank you for your telephone call yesterday. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

I would like to reassure you that this antibody is tested and covered by our 6 month guarantee for ELI...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - nuclear (Hela)
Loading amount
10 µg
Specification
Hela
Treatment
100 µM CoCl2 15hrs
Gel Running Conditions
Reduced Denaturing (8% gel)
Blocking step
Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 21°C
Username

Dr. Sridhar Vasudevan

Verified customer

Submitted Jul 23 2012

Thank you for your enquiry.

I can confirm that the anti-HIF1 alpha antibody [1A3] ab113642is sold as ascites supernatant. Unpurified antibodies, such as those sold as whole anti-serum, ascites or tissue culture supernatant will not have a co...

Read More

Thank you for contacting us yesterday.
As discussed over the phone, I have looked into whether there might be any suitable alternative to the Goat anti-Mouse IgG H&L (Chromeo™ 546) secondary antibody (ab60316) for you. Particularly in regards to s...

Read More

Thank you for your reply. I am sorry we have no other HIF1 alpha antibodies that are tested in ChIP. We do have several tested in WB and IHC-P. I have checked the alignment of the immunogen of these antibodies with the zebrafish sequence. I am sorry th...

Read More

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