Anti-HIF-1-alpha antibody - ChIP Grade (ab2185)

Overview

  • Product nameAnti-HIF-1-alpha antibody - ChIP GradeSee all HIF-1-alpha primary antibodies ...
  • Description
    Rabbit polyclonal to HIF-1-alpha - ChIP Grade
  • Tested applicationsIHC-P, WB, IP, Gel supershift assays, IHC-Fr, ICC, ICC/IF, ChIPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Guinea pig, Human, Xenopus laevis, Monkey
    Predicted to work with: Cow
  • Immunogen

    Fusion protein corresponding to Human HIF-1-alpha aa 432-528.

Properties

Applications

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.

Application Abreviews Notes
IHC-P 1/100.
WB 1/500 - 1/1000.
IP 1/1000.
Gel supershift assays 1/1 - 1/100.
IHC-Fr 1/10 - 1/2000.
ICC Use at an assay dependent concentration.
ICC/IF 1/10 - 1/2000.
ChIP Use 25µl for 106 cells.

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
    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 localizationCytoplasm. 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 - ChIP Grade images

  • 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human kidney tissue sections labeling HIF-1-alpha with ab2185.

  • All lanes : Anti-HIF-1-alpha antibody - ChIP Grade (ab2185)

    Lane 1 : Rat nuclear extract lysate - normoxic
    Lane 2 : Rat nuclear extract lysate - hypoxic
  • ChIP analysis of HIF-1-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.

References for Anti-HIF-1-alpha antibody - ChIP Grade (ab2185)

This product has been referenced in:
  • Zheng X  et al. Hypoxia-induced and calpain-dependent cleavage of filamin A regulates the hypoxic response. Proc Natl Acad Sci U S A 111:2560-5 (2014). ChIP . Read more (PubMed: 24550283) »
  • Cheverud JM  et al. Fine-mapping quantitative trait loci affecting murine external ear tissue regeneration in the LG/J by SM/J advanced intercross line. Heredity (Edinb) 112:508-18 (2014). Mouse . Read more (PubMed: 24569637) »

See all 29 Publications for this product

Product Wall

Abreviews
Application Immunocytochemistry
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Sample Mouse Cultured Cells (Neuro2A- neuroblastoma)
Specification Neuro2A- neuroblastoma
Permeabilization Yes - 0.3% Triton X-100
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Jun 04 2013

Thank you for contacting us.

ab2185 Immunogen sequence is more than 90% similar with Pig HIF1 alpha so this antibody is more likely to cross react.

http://www.ebi.ac.uk/Tools/services/web_clustalw2/toolresult.ebi?tool=clustalw2&jo...

Read More
Application Western blot
Sample Mouse Tissue lysate - whole (Striatum (Brain))
Loading amount 50 µg
Specification Striatum (Brain)
Gel Running Conditions Reduced Denaturing (8%)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Feb 14 2013

Thank you for your reply. The immunogen shows 89% homology with the immunogen. We are not able to obtain consistent results with rat samples therefore are not able to guarantee that it will give positive results. However, customers have shown that t...

Read More

Thank you for sending the image.

I now think that the bands are an indication of no band. If no protein can be bound antibodies often produce a lot of background.

This background is also often generated by the secondary antibody. Read More

Thank you and your customer for taking the time to complete our questionnaire and contact us. I am sorry to hear your customer has had difficulty obtaining satisfactory results from this antibody.

The details your customer has kindly provided...

Read More

Thank you for your reply.

I am sorry that ab51608 on original order xxxxxx did not work and we will add a new vial of ab2185 to your next order.

Please accept my apologies for the problems with this antibody and do not hesitate to con...

Read More

Thank you for taking the time send us this information. Your enquiry has been forwarded to me as my colleague ### is currently away from the office. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.
Read More

Thank you for contacting us on Friday.

As discussed over the phone, unfortunately the anti-HIF1 alpha antibody ab2185 which you were hoping to use in ChIP is not likely to be available until the end of January. The only other antibody which we...

Read More

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

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"