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.
A novel putative proline hydroxylase, PH-4, with an N-terminal EF-hand motif and a C-terminal catalytic domain with 1 transmembrane domain has been identified in endoplasmic reticulum.
PH-4 showed approx.
10-15% homology with the collagen prolyl 4-hydroxylases, PH-alpha I, and PH-alpha II.
Human PH-4 (502aa, chromosome 3pp21.3) is highly expressed in most tissues.
Like PHD1-3, PH-4 suppressed the HIF transactivation activity, dependent on the consensus oxygen-dependent degradation domain (ODDD) proline residues.
Ph-4 levels correlated with the cellular HIF concentration.
Therefore, PH-4 may also be involved in cellular oxygen sensing mechanism.
Applications:Suitable for use in Western Blot and ELISA.
Other applications not tested.
Recommended Dilution:Western Blot: 1-5ug/ml (chemiluminescence)ELISA: 1:10,000-1:100,000; 50-100ng control peptide/wellOptimal 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.
仕様
Size：100ul
Host：rabbit
Source Antibody：human
Grade：Affinity Purified
Purity：Purified by immunoaffinity chromatography.
Form：Supplied as a lyophilized powder from PBS, 0.05% sodium azide.
Specificity：Recognizes human PH-4.
Isotype：IgG
Calc Applications Abbrev：E WB
Calc Crossreactivity：Hu
Immunogen：Synthetic peptide corresponding to 17aa of human PH-4, C-terminal.
