Technical Data
PHD3 (Prolyl Hydroxylase Domain Containing Protein 3, HIF-Prolyl Hydroxylase 3, EGLN3)
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. HPH-1, 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.

Suitable for use in Western Blot., Immunofluorescence, Immunocytochemistry, Immunohistochemistry, Immunoprecipitation and Electron Microscopy. Other applications not tested.

Recommended Dilutions:
Western Blot: 1:1000-1:2000
Immunofluorescence (IC): 1:500
Immunohistochemistry: Paraffin sections
Optimal dilution to be determined by the researcher.

Positive Control:
HeLa cell lysates

Storage and Stability:
May be stored at 4C. For long-term storage, aliquot and store at 4C. Do not freeze. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial prior to removing the cap. Further dilutions can be made in assay buffer.
PabIgGAffinity Purified
100ul4C Do not freezeBlue IceHumanRabbit
A synthetic peptide corresponding to a region between residues 50-100 of human PHD3 (NP_071356.1, GeneID 112399), Species Sequence Homology: mouse
Purified by immunoaffinity chromatography
Supplied as a liquid in Tris-citrate/phosphate, pH 7.5, 0.09% sodium azide
Recognizes human PHD3 at ~28kD. Species Crossreactivity: rat
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. Bruick, RK, McKnight, SL. A Conserved Family of Prolyl-4-Hydroxylases That Modify HIF. Science. 294:1337-1340(2001). 2. Ivan, M, et al. Biochemical Purification and Pharmacological Inhibition of a Mammalian Prolyl Hydroxylase Acting on Hypoxia-Inducible Factor. PNAS. 99(21):13459-13464(2002). 3. Maxwell, PH, Ratcliffe, PJ. Oxygen Sensors and Angiogenesis. Cell & Dev. Bio. 13:29-37(2002).