Technical Data

P3374-05
Clone Type
Polyclonal
Host
Rabbit
Source
Human
Isotype
IgG
Grade
Affinity Purified
Applications
E
Crossreactivity
Hu
Shipping Temp
Blue Ice
Storage Temp
-20°C
Rabbit Anti-PHD2 (PH2, Prolyl hydroxylase domain containing protein 2, HIF2PH2, HIF-Prolyl hydroxylase 2, EGLN1, egl nine homolog 1, C1orf12, chromosome 1 open reading frame 12)

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. PHD2 (human 426 aa, chromosome 1q42-q43, also known as Egl nine homolog 1, EGLN1, (Hypoxia-inducible factor prolyl hydroxylase 2) (HIF-prolyl hydroxylase 2) (HIF-PH2) (HPH-2) (Prolyl hydroxylase domain-containing protein 2) PHD2 (SM-20) (PNAS-118 / PNAS-137) is widely expressed. Alternatively spliced isoforms 2 is missing 337-358 aa. It is activated by hypoxia in some cells and tissues.

Applications
Suitable for use in ELISA. Western Blot, though not tested, may potentially be used as an application. Other applications not tested.
Recommended Dilution
ELISA: 1:10,000-1:100,000; Using 50-100ng of control peptide/well. See P3374-05A. Western Blot: 1-5ug/ml using ECL technique. Optimal dilutions to be determined by the researcher.
Storage and Stability
Lyophilized powder may be stored at -20°C. Stable for 12 months at -20°C. Reconstitute with sterile ddH2O or PBS. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Reconstituted product is stable for 12 months at -20°C. 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.
Immunogen
16aa peptide of human PHD2/EGLN1. Location: ~C-terminal. Species Sequence Homology: Chicken - 87%; Mouse, rat - 68%.
Form
Supplied as a lyophilized powder from PBS, 0.05% sodium azide.
Purity
Purified by immunoaffinity chromatography.
Specificity
Recognizes human PHD2.

Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.

References
Dupuy D et al (2000) Genomincs 69, 348-354; Taylor MS et al (2001) Gene 275, 125-132; Semenza GL et al (2001) Cell, 107, 1-3; Epstein ACR et al (2001) Cell 107, 43-54; Ivan M et al (2002) PNAS 99, 13459-13464; Cioffi CL et al BBRC 303, 947-953
USBio References
No references available
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