Recombinant Human HIF-1 alpha protein (ab154478)

Overview

Description

  • Nature
    Recombinant
  • Source
    Escherichia coli
  • Amino Acid Sequence
    • Accession
    • Species
      Human
    • Sequence
      MSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDEIADEY QGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQL KEFLDANLAGSGSGHMHHHHHHSSGLVPRGSGMKETAAAKFERQHMDSPD LGTENLYFQGMEGAGGANDKKKISSERRKEKSRDAARSRRSKESEVFYEL AHQLPLPHNVSSHLDKASVMRLTISYLRVRKLLDAGDLDIEDDMKAQMNC FYLKALDGFVMVLTDDGDMIYISDNVNKYMGLTQFELTGHSVFDFTHPCD HEEMREMLTHRNGLVKKGKEQNTQRSFFLRMKCTLTSRGRTMNIKSATWK VLHCTGHIHVYDTNSNQPQCGYKKPPMTCLVLICEPIPHPSNIEIPLDSK TFLSRHSLDMKFSYCDERITELMGYEPEELLGRSIYEYYHALDSDHLTKT HHDMFTKGQVTTGQYRMLAKRGGYVWVETQATVIYNTKNSQPQCIVCVNY VVSGIIQHDLIFSLQQTECVLKPVESSDMKMTQLFTKVESEDTSSLFDKL KKEPDALTLLAPAAGDTIISLDFGSNDTETDDQQLEEVPLYNDVMLPSPN EKLQNINLAMSPLPTAETPKPLRSSADPALNQEVALKLEPNPESLELSFT MPQIQDQTPSPSDGSTRQSSPEPNSPSEYCFYVDSDMVNEFKLELVEKLF AEDTEAKNPFSTQDTDLDLEMLAPYIPMDDDFQLRSFDQLSPLESSSASP ESASPQSTVTVFQQTQIQEPTANATTTTATTDELKTVTKDRMEDIKILIA SPSPTHIHKETTSATSSPYRDTQSRTASPNRAGKGVIEQTEKSHPRSPNV LSVALSQRTTVPEEELNPKILALQNAQRKRKMEHDGSLFQAVGIG
    • Molecular weight
      100 kDa including tags
    • Amino acids
      1 to 735
    • Tags
      His tag N-Terminus

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.

  • Applications

    ELISA

    Western blot

    SDS-PAGE

  • Purity
    > 75 % SDS-PAGE.
    ab154478 was purified by Ni chromatography and sterile filtered.
  • Form
    Liquid
  • Additional notes

    Product was previously marketed under the MitoSciences sub-brand.

  • Concentration information loading...

Preparation and Storage

  • Stability and Storage

    Shipped on dry ice. Upon delivery aliquot and store at -80ºC. Avoid freeze / thaw cycles.

    pH: 8.00
    Constituents: 0.61% Tris, 10% Glycerol, 0.88% Sodium chloride

General Info

  • Alternative names
    • ARNT interacting protein
    • ARNT-interacting protein
    • Basic helix loop helix PAS protein MOP1
    • Basic-helix-loop-helix-PAS protein MOP1
    • bHLHe78
    • Class E basic helix-loop-helix protein 78
    • HIF 1A
    • HIF 1alpha
    • HIF-1-alpha
    • HIF1
    • HIF1 A
    • HIF1 Alpha
    • HIF1-alpha
    • HIF1A
    • HIF1A_HUMAN
    • Hypoxia inducible factor 1 alpha
    • Hypoxia inducible factor 1 alpha isoform I.3
    • Hypoxia inducible factor 1 alpha subunit
    • Hypoxia inducible factor 1 alpha subunit basic helix loop helix transcription factor
    • Hypoxia inducible factor 1, alpha subunit (basic helix loop helix transcription factor)
    • Hypoxia inducible factor1alpha
    • Hypoxia-inducible factor 1-alpha
    • Member of PAS protein 1
    • Member of PAS superfamily 1
    • Member of the PAS Superfamily 1
    • MOP 1
    • MOP1
    • PAS domain-containing protein 8
    • PASD 8
    • PASD8
    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
    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

Images

  • 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.

  • 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

  • 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.

References

ab154478 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

HIF1alpha is a large protein to express recombinantly in E.coli so a few elements were added to aid in expression/solubility/purification. This construct has an N-terminal Trx – His Tag followed by a linker and a TEV cleavage site.

-Tr...

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