Technical Data
P3375-06
PHD3 (HIF-Prolyl hydroxylase 3, EGLN3)
Description:
HIF prolyl hydroxylase 3 is a prolyl hydroxylase that modifies HIF-alpha. Classic prolyl hydroxylases are found in the endoplasmic reticulum and modify collagen, whereas HIF is an intracellular protein and the HPH sites do not resemble those modifying collagen. HIF is a transcriptional complex that plays a critical role in oxygen homeostasis. HPH is an essential component of the pathway through which cells sense oxygen. In the presence of oxygen, HPHs convert specific prolyl residues in HIF-alpha to hydroxyproline, leading to HIF-alpha destruction. Low oxygen levels, sensed at the cellular level, cause the HIF conversion to be reduced so that HIF is stable and there is increased angiogenesis.
HIF prolyl hydroxylase 3, specifically, catalyzes the post-translational formation of 4-hydroxyproline in HIF alpha proteins. It hydroxylates HIF-1 alpha at Pro(564) and HIF-2 alpha. It targets HIF through the hydroxylation for proteasomal degradation via the von Hippel-Lindau ubiquitylation complex. It may also play a role in cell growth regulation in muscle cells and in apoptosis in neuronal tissues, promoting cell death through a caspase-dependent mechanism.

Applications:
Suitable for use in Western Blot. Other applications not tested.

Recommended Dilution:
Western Blot: 1:1000-1:3000 (ECL). A band is seen at ~27kD, representing HIF prolyl hydroxylase 3. Has been tested against HeLa and MEF cell lysates.
Optimal dilutions to be determined by the researcher.

Storage and Stability:
May be stored at 4C for short-term only. For long-term storage and to avoid repeated freezing and thawing, add sterile glycerol (40-50%), aliquot and store at -20C. Aliquots are stable for at least 12 months at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
TypeIsotypeCloneGrade
PabIgGAffinity Purified
SizeStorageShippingSourceHost
100ul4C (-20C Glycerol)Blue IceHumanRabbit
Concentration:
~1mg/ml
Immunogen:
A synthetic peptide made to the C-terminus of the human protein sequence of HIF prolyl hydroxylase 3 [LocusLink ID 112399].
Purity:
Purified by Protein G affinity chromatography.
Form
Supplied as a liquid in Tris-citrate buffer with 0.01% sodium azide.
Specificity:
Specific for human HIF prolyl hydroxylase 3. Species Crossreactivity: Mouse. Reactivity is likely with rat due to 26/28 residue homology between the rat protein and the peptide used as the immunogen for this antibody.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. Bruick, R.K. & McKnight, S.L., Science 294: 1337-1340 (2001). 2. Ivan, M., et al., PNAS 99(21): 13,459-13,464 (2002). 3. Maxwell, P.H. & Ratcliffe, P.J., Cell & Dev. Bio. 13: 29-37 (2002).