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Recombinant PHD2 (EGLN1) protein

Expressed In

Baculovirus

Protein Species

Human

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Cat No : 81065

Validation Data Gallery

  • Recombinant PHD2 (EGLN1) protein gel 10% SDS-PAGE gel with Coomassie blue staining MW: 47 kDa Purity: > 90%

    Recombinant PHD2 (EGLN1) protein gel 10% SDS-PAGE gel with Coomassie blue staining MW: 47 kDa Purity: > 90%

  • MALDI-TOF for PHD2 (EGLN1) protein activity 3 uM HIF1A (HIF-1a) peptide (peptide sequence: biotin-DLDLEALAPYIPADDDFQL) was incubated with 300 nM PHD2 (EGLN1) protein in a 30 ul reaction system containing 20 mM Tris-HCl pH 7.5, 5 mM KCl, 1.5 mM MgCl2, 1 mM DTT, 100 uM 2-oxoglutarate, 100 uM ascorbate and 50 uM (NH4)2Fe(SO4)2·6H2O for 2 hr at 3&degree;. Single 3 uM HIF1A peptide was used as negative control. MALDI-TOF was used for detection.

    MALDI-TOF for PHD2 (EGLN1) protein activity 3 uM HIF1A (HIF-1a) peptide (peptide sequence: biotin-DLDLEALAPYIPADDDFQL) was incubated with 300 nM PHD2 (EGLN1) protein in a 30 ul reaction system containing 20 mM Tris-HCl pH 7.5, 5 mM KCl, 1.5 mM MgCl2, 1 mM DTT, 100 uM 2-oxoglutarate, 100 uM ascorbate and 50 uM (NH4)2Fe(SO4)2·6H2O for 2 hr at 3°ree;. Single 3 uM HIF1A peptide was used as negative control. MALDI-TOF was used for detection.

Product Information

Expressed In Baculovirus
Protein Species Human

Contents

Recombinant PHD2 / EGLN1 protein is supplied in 25 mM HEPES-NaOH pH 7.5, 300 mM NaCl, 10% glycerol, 0.04% Triton X-100 and 0.5 mM TCEP.

Background

Prolyl Hydroxylase Domain-Containing Protein 2 (PHD2) or Egl-9 Family Hypoxia Inducible Factor 1 (EGLN1), also known as HIF-PH2 or HPH-2, is a prolyl hydroxylase. PHD3 is a cellular oxygen sensor that catalyzes the post-translational formation of 4-hydroxyproline in hypoxia-inducible factor (HIF) alpha proteins under normoxic conditions. It can hydroxylate a specific proline found in each of the oxygen-dependent degradation (ODD) domains (N-terminal, NODD, and C-terminal, CODD) of HIF1A, also HIF2A. PHD1 has a preference for the CODD site for both HIF1A and HIF2A. Hydroxylated HIFs are targeted for proteasomal degradation via the von Hippel-Lindau ubiquitination complex. Under hypoxic conditions, the hydroxylation reaction is attenuated allowing HIFs to escape degradation resulting in their translocation to the nucleus, heterodimerization with HIF1B, and increased expression of hypoxy-inducible genes. PHD2 is the most important isozyme under normoxia and, through regulating the stability of HIF1, involved in various hypoxia-influenced processes such as angiogenesis in retinal and cardiac functionality. Its target proteins are preferentially recognized via a LXXLAP motif.

Application Notes

This product was manufactured as described in Protein Details. Where possible, Active Motif has developed functional or activity assays for recombinant proteins. Additional characterization such as enzyme kinetic activity assays, inhibitor screening or other biological activity assays may not have been performed for every product. All available data for a given product is shown on the lot-specific Technical Data Sheet.

Protein Details

Recombinant human PHD2 (EGLN1) protein was expressed in a baculovirus expression system as the full length protein (accession number NP_071334.1) with an N-terminal FLAG-Tag. The molecular weight of the protein is 47 kDa.

Storage

Recombinant proteins in solution are temperature sensitive and must be stored at -80°C to prevent degradation. Avoid repeated freeze/thaw cycles and keep on ice when not in storage.

Guarantee

This product is guaranteed for 6 months from date of receipt.

This product is for research use only and is not for use in diagnostic procedures.