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., Immunohistochemistry, Immunofluorescence/Immunocytochemistry, Immunoprecipitation and Electron Microscopy. Other applications not tested.

Recommended Dilutions:
Western Blot: 1:1000-1:2000
Immunofluorescence (IC): 1:500
Immunohistochemistry (Paraffin): 1:250-1:1000
Optimal dilution to be determined by the researcher.

Positive Control:
HeLa cell lysates

Storage and Stability:
May be stored at 4C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
PabIgGAffinity Purified
100ul-20CBlue 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 PBS, pH 7.5, 0.05% 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).