Recombinant Human HIF-1 alpha protein (ab154478)
Key features and details
- Expression system: Escherichia coli
- Purity: > 75% SDS-PAGE
- Tags: His tag N-Terminus
- Suitable for: ELISA, WB, SDS-PAGE
Description
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Product name
Recombinant Human HIF-1 alpha protein
See all HIF-1 alpha proteins and peptides -
Purity
> 75 % SDS-PAGE.
ab154478 was purified by Ni chromatography and sterile filtered. -
Expression system
Escherichia coli -
Accession
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Protein length
Full length protein -
Animal free
No -
Nature
Recombinant -
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Species
Human -
Sequence
MSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDEIADEY QGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQL KEFLDANLAGSGSGHMHHHHHHSSGLVPRGSGMKETAAAKFERQHMDSPD LGTENLYFQGMEGAGGANDKKKISSERRKEKSRDAARSRRSKESEVFYEL AHQLPLPHNVSSHLDKASVMRLTISYLRVRKLLDAGDLDIEDDMKAQMNC FYLKALDGFVMVLTDDGDMIYISDNVNKYMGLTQFELTGHSVFDFTHPCD HEEMREMLTHRNGLVKKGKEQNTQRSFFLRMKCTLTSRGRTMNIKSATWK VLHCTGHIHVYDTNSNQPQCGYKKPPMTCLVLICEPIPHPSNIEIPLDSK TFLSRHSLDMKFSYCDERITELMGYEPEELLGRSIYEYYHALDSDHLTKT HHDMFTKGQVTTGQYRMLAKRGGYVWVETQATVIYNTKNSQPQCIVCVNY VVSGIIQHDLIFSLQQTECVLKPVESSDMKMTQLFTKVESEDTSSLFDKL KKEPDALTLLAPAAGDTIISLDFGSNDTETDDQQLEEVPLYNDVMLPSPN EKLQNINLAMSPLPTAETPKPLRSSADPALNQEVALKLEPNPESLELSFT MPQIQDQTPSPSDGSTRQSSPEPNSPSEYCFYVDSDMVNEFKLELVEKLF AEDTEAKNPFSTQDTDLDLEMLAPYIPMDDDFQLRSFDQLSPLESSSASP ESASPQSTVTVFQQTQIQEPTANATTTTATTDELKTVTKDRMEDIKILIA SPSPTHIHKETTSATSSPYRDTQSRTASPNRAGKGVIEQTEKSHPRSPNV LSVALSQRTTVPEEELNPKILALQNAQRKRKMEHDGSLFQAVGIG -
Predicted molecular weight
100 kDa including tags -
Amino acids
1 to 735 -
Tags
His tag N-Terminus
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Associated products
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Related Products
Specifications
Our Abpromise guarantee covers the use of ab154478 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
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Applications
ELISA
Western blot
SDS-PAGE
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Form
Liquid -
Additional notes
Product was previously marketed under the MitoSciences sub-brand.
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Concentration information loading...
Preparation and Storage
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Stability and Storage
Shipped on Dry Ice. Store at -80°C. Avoid freeze / thaw cycle.
pH: 8.00
Constituents: 0.61% Tris, 10% Glycerol, 0.88% Sodium chloride
General Info
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Alternative names
- ARNT interacting protein
- ARNT-interacting protein
- Basic helix loop helix PAS protein MOP1
see all -
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
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 localization
Cytoplasm. Nucleus. Cytoplasmic in normoxia, nuclear translocation in response to hypoxia. Colocalizes with SUMO1 in the nucleus, under hypoxia. - Information by UniProt
Images
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SDS-PAGE - Recombinant Human HIF-1 alpha protein (ab154478)
1µg of ab154478 was examined by SDS-PAGE and commassie blue protein stain. Full-length HIF-1-alpha is indicated by the arrow. Purity is judged to be >75%.
Expected MW is 100kDa.
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Western blot - Recombinant Human HIF-1 alpha protein (ab154478)ab154478 was examined by western blot with an anti-HIF-1-alpha antibody.
Lane1: 10ng HIF1 alpha full-length protein (ab154478)
Block: 4% milk/PBS Primary antibody: anti-HIF-1-alpha (ab51608), 1:1000; 2 hours room temperature. Secondary antibody: anti-Rabbit HRP 1:5000 ECL detection
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ELISA - Recombinant Human HIF-1 alpha protein (ab154478)ab154478 was tested in the HIF1A Human ELISA Kit (ab117996). ab154478 was tested under standard conditions in the sandwich ELISA kit using a 3-fold dilution series from 3µg/ml.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (4)
ab154478 has been referenced in 4 publications.
- Dawn Woodfield J et al. Synthesis, Binding Affinity Analysis, and 18 F Radiosynthesis of Small-Molecular-Weight HIF-1α-Binding Compounds. ChemMedChem 17:e202100544 (2022). PubMed: 34595843
- Sil S et al. HIV-1 Tat-mediated astrocytic amyloidosis involves the HIF-1a/lncRNA BACE1-AS axis. PLoS Biol 18:e3000660 (2020). PubMed: 32453744
- Duda P et al. The Reverse Warburg Effect is Associated with Fbp2-Dependent Hif1a Regulation in Cancer Cells Stimulated by Fibroblasts. Cells 9:N/A (2020). PubMed: 31947613
- Chen X et al. Hypoxia inducible factor and microvessels in peri-implantation endometrium of women with recurrent miscarriage. Fertil Steril 105:1496-1502.e4 (2016). PubMed: 27018158