Technical Data
P3373A
PHD1-3, pan (HIF-prolyl Hydroxylase 1-3)
Description:
Oxygen is absolutely critical for the survival of mammalian cells. Hypoxia induced factor (HIF) is a transcriptional complex that plays a central role in mammalian oxygen homeostasis. There are 3-types of alpha subunits (HIF1-3alpha) and one HIF-beta subunits. However, the three HIF-alpha subunits are regulated by oxygen in a similar fashion, i.e. by regulated stabilization of the alpha-subunits. Under normal conditions, HIF-alpha Prolines are hydroxylated at Pro-402 and Pro-564. This allows binding of von Hippel-Lindau (VHL), the substrate recognition component of the E3 ubiquinated ligase complex, subsequent ubiquitination and degradation of HIF-alpha by the proteasome. Under hypoxic conditions, hydroxylation of HIF-alpha is inhibited and this prevents HIF-alpha degradation. The enzymes responsible for HIF-hydroxylation are known as HIF-prolyl hydroxylases (PHD1-3 or HPH1, HPH2, and HPH3). The three PHDs have been identified to hydroxylate the motif, LXXLAP* with *P being the hydroxyproline.

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

Recommended Dilution:
ELISA: 1:10,000-1:100,000; Using 50-100ng of P3373B control peptide/well.
Western Blot: 1-5ug/ml. Using ECL technique.
Optimal dilutions to be determined by the researcher.

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 at least 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
TypeIsotypeCloneGrade
PabIgGAffinity Purified
SizeStorageShippingSourceHost
50ug-20CBlue IceHumanRabbit
Concentration:
~1mg/ml
Immunogen:
Synthetic peptide corresponding to 18 amino acids near the N terminus of human PHD2/EGLN1.
Purity:
Purified by immunoaffinity chromatography.
Form
Supplied as a liquid in PBS, pH 7.2, 0.1% BSA.
Specificity:
Recognizes human PHD1-3. Species Sequence Homology: mouse, rat, PHD2/EGLN1-100%; human, mouse, rat PHD1/EGLN2, 94%; human, mouse, rat PHD3/EGLN3, 88%.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. trausberg RL et al (2002) PNAS 99, 16899-16903; Taylor MS et al (2001) Gene 275, 125-132; 2. Epstein AC et al (2001) Cell 107, 43-54; Semenza GL et al (2001) Cell 107, 1-3; 3. Bruick RK et al (2001) Science 294, 1337-1340; Oehme F et al (2002) BBRC 296, 343-349