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Immunofluorescence (IF)/Immunocytochemistry (ICC) (staining Hela cells by IF/ICC. The samples were fixed with PFA and permeabilized in 0.1% Triton X-100,then blocked in 10% serum for 45 minutes at 25 degree C. Samples were then incubated with primary Ab(1:200) and mouse anti-beta tubulin Ab(1:200) for 1 hour at 37 degree C. An AlexaFluor594 conjugated goat anti-rabbit IgG(H+L) Ab(1:200 Red) and an AlexaFluor488 conjugated goat anti-mouse IgG(H+L) Ab(1:600 Green) were used as the secondary antibod)

Rabbit PHD3 Polyclonal Antibody | anti-PHD3 antibody

PHD3 Antibody

Gene Names
EGLN3; PHD3; HIFPH3; HIFP4H3
Reactivity
Human, Mouse
Predicted: Pig, Bovine, Horse, Sheep, Rabbit
Applications
Western Blot
Purity
The antiserum was purified by peptide affinity chromatography using SulfoLink Coupling Resin (Thermo Fisher Scientific).
Synonyms
PHD3; Polyclonal Antibody; PHD3 Antibody; Egl 9 family hypoxia inducible factor 3; Egl nine homolog 3 (C. elegans); Egl nine homolog 3; Egl nine like protein 3 isoform; EGL9 homolog of C. elegans 3; EGLN3; EGLN3_HUMAN; Factor responsive smooth muscle protein; HIF Prolyl Hydroxylase 3; HIF-PH3; HIF-prolyl hydroxylase 3; HIFP4H3; HIFPH3; HPH-1; HPH-3; Hypoxia-inducible factor prolyl hydroxylase 3; P4H3; Prolyl Hydroxylase Domain Containing Protein 3; Prolyl hydroxylase domain-containing protein 3; SM20; anti-PHD3 antibody
Ordering
For Research Use Only!
Host
Rabbit
Reactivity
Human, Mouse
Predicted: Pig, Bovine, Horse, Sheep, Rabbit
Clonality
Polyclonal
Isotype
IgG
Specificity
PHD3 Antibody detects endogenous levels of PHD3.
Purity/Purification
The antiserum was purified by peptide affinity chromatography using SulfoLink Coupling Resin (Thermo Fisher Scientific).
Form/Format
Liquid; Phosphate buffered saline, pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.
Concentration
1mg/ml (varies by lot)
Sequence Length
239
Applicable Applications for anti-PHD3 antibody
Western Blot (WB)
Application Notes
WB: 1:500-1:2000
Immunogen
A synthesized peptide derived from human PHD3.
Tissue Specificity
Widely expressed at low levels. Expressed at higher levels in adult heart (cardiac myocytes, aortic endothelial cells and coronary artery smooth muscle), lung and placenta, and in fetal spleen, heart and skeletal muscle. Also expressed in pancreas. Localized to pancreatic acini and islet cells.
Fragment
Fab Fragment
Conjugate
Unconjugated
Subcellular Location
Nucleus
Preparation and Storage
Store at -20 degree C. Stable for 12 months from date of receipt.

Immunofluorescence (IF)/Immunocytochemistry (ICC)

(staining Hela cells by IF/ICC. The samples were fixed with PFA and permeabilized in 0.1% Triton X-100,then blocked in 10% serum for 45 minutes at 25 degree C. Samples were then incubated with primary Ab(1:200) and mouse anti-beta tubulin Ab(1:200) for 1 hour at 37 degree C. An AlexaFluor594 conjugated goat anti-rabbit IgG(H+L) Ab(1:200 Red) and an AlexaFluor488 conjugated goat anti-mouse IgG(H+L) Ab(1:600 Green) were used as the secondary antibod)

Immunofluorescence (IF)/Immunocytochemistry (ICC) (staining Hela cells by IF/ICC. The samples were fixed with PFA and permeabilized in 0.1% Triton X-100,then blocked in 10% serum for 45 minutes at 25 degree C. Samples were then incubated with primary Ab(1:200) and mouse anti-beta tubulin Ab(1:200) for 1 hour at 37 degree C. An AlexaFluor594 conjugated goat anti-rabbit IgG(H+L) Ab(1:200 Red) and an AlexaFluor488 conjugated goat anti-mouse IgG(H+L) Ab(1:600 Green) were used as the secondary antibod)

Western Blot (WB)

(Western blot analysis of extracts from B16F10, using PHD3 Antibody. The lane on the left was treated with blocking peptide.Observed bands: 20 kDa.)

Western Blot (WB) (Western blot analysis of extracts from B16F10, using PHD3 Antibody. The lane on the left was treated with blocking peptide.Observed bands: 20 kDa.)

Western Blot (WB)

(Western blot analysis of extracts from HEK-293 cells, using PHD3 Antibody. The lane on the left was treated with blocking peptide.)

Western Blot (WB) (Western blot analysis of extracts from HEK-293 cells, using PHD3 Antibody. The lane on the left was treated with blocking peptide.)
Related Product Information for anti-PHD3 antibody
Description: Cellular oxygen sensor that catalyzes, under normoxic conditions, the post-translational formation of 4-hydroxyproline in hypoxia-inducible factor (HIF) alpha proteins. Hydroxylates a specific proline found in each of the oxygen-dependent degradation (ODD) domains (N-terminal, NODD, and C-terminal, CODD) of HIF1A. Also hydroxylates HIF2A. Has a preference for the CODD site for both HIF1A and HIF2A. Hydroxylation on the NODD site by EGLN3 appears to require prior hydroxylation on the CODD site. Hydroxylated HIFs are then targeted for proteasomal degradation via the von Hippel-Lindau ubiquitination complex. Under hypoxic conditions, the hydroxylation reaction is attenuated allowing HIFs to escape degradation resulting in their translocation to the nucleus, heterodimerization with HIF1B, and increased expression of hypoxy-inducible genes. EGLN3 is the most important isozyme in limiting physiological activation of HIFs (particularly HIF2A) in hypoxia. Also hydroxylates PKM in hypoxia, limiting glycolysis. Under normoxia, hydroxylates and regulates the stability of ADRB2. Regulator of cardiomyocyte and neuronal apoptosis. In cardiomyocytes, inhibits the anti-apoptotic effect of BCL2 by disrupting the BAX-BCL2 complex. In neurons, has a NGF-induced proapoptotic effect, probably through regulating CASP3 activity. Also essential for hypoxic regulation of neutrophilic inflammation. Plays a crucial role in DNA damage response (DDR) by hydroxylating TELO2, promoting its interaction with ATR which is required for activation of the ATR/CHK1/p53 pathway. Target proteins are preferentially recognized via a LXXLAP motif.

Subunit Structure: Interacts with WDR83; the interaction leads to almost complete elimination of HIF-mediated reporter activity (By similarity). Interacts with BCL2 (via its BH4 domain); the interaction disrupts the BAX-BCL4 complex inhibiting the anti-apoptotic activity of BCL2. Interacts with ADRB2; the interaction hydroxylates ADRB2 facilitating its ubiquitination by the VHL-E3 ligase complex. Interacts with PAX2; the interaction targets PAX2 for destruction. Interacts with PKM; the interaction hydroxylates PKM in hypoxia.

Similarity: The Beta2beta3 'finger-like' loop domain is important for substrate (HIFs' CODD/NODD) selectivity.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
UniProt Accession #
Molecular Weight
Observed: 28 kDa
Predicted: 28 kDa
NCBI Official Full Name
Egl nine homolog 3
NCBI Official Synonym Full Names
egl-9 family hypoxia inducible factor 3
NCBI Official Symbol
EGLN3
NCBI Official Synonym Symbols
PHD3; HIFPH3; HIFP4H3
NCBI Protein Information
egl nine homolog 3
UniProt Protein Name
Egl nine homolog 3
UniProt Gene Name
EGLN3
UniProt Synonym Gene Names
HIF-PH3; HIF-prolyl hydroxylase 3; HPH-3; PHD3
UniProt Entry Name
EGLN3_HUMAN

Uniprot Description

EGLN3: Cellular oxygen sensor that catalyzes, under normoxic conditions, the post-translational formation of 4-hydroxyproline in hypoxia-inducible factor (HIF) alpha proteins. Hydroxylates a specific proline found in each of the oxygen-dependent degradation (ODD) domains (N-terminal, NODD, and C-terminal, CODD) of HIF1A. Also hydroxylates HIF2A. Has a preference for the CODD site for both HIF1A and HIF2A. Hydroxylation on the NODD site by EGLN3 appears to require prior hydroxylation on the CODD site. Hydroxylated HIFs are then targeted for proteasomal degradation via the von Hippel-Lindau ubiquitination complex. Under hypoxic conditions, the hydroxylation reaction is attenuated allowing HIFs to escape degradation resulting in their translocation to the nucleus, heterodimerization with HIF1B, and increased expression of hypoxy-inducible genes. EGLN3 is the most important isozyme in limiting physiological activation of HIFs (particularly HIF2A) in hypoxia. Also hydroxylates PKM in hypoxia, limiting glycolysis. Under normoxia, hydroxylates and regulates the stability of ADRB2. Regulator of cardiomyocyte and neuronal apoptosis. In cardiomyocytes, inhibits the anti-apoptotic effect of BCL2 by disrupting the BAX-BCL2 complex. In neurons, has a NGF-induced proapoptotic effect, probably through regulating CASP3 activity. Also essential for hypoxic regulation of neutrophilic inflammation.

Protein type: Oxidoreductase; EC 1.14.11.29

Chromosomal Location of Human Ortholog: 14q13.1

Cellular Component: nucleoplasm; cytoplasm; nucleus; cytosol

Molecular Function: protein binding; L-ascorbic acid binding; iron ion binding; oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, 2-oxoglutarate as one donor, and incorporation of one atom each of oxygen into both donors; peptidyl-proline 4-dioxygenase activity

Biological Process: caspase activation; apoptosis; regulation of neuron apoptosis; response to hypoxia; protein amino acid hydroxylation; peptidyl-proline hydroxylation to 4-hydroxy-L-proline; response to DNA damage stimulus; regulation of cell proliferation

Research Articles on PHD3

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Product Notes

The PHD3 egln3 (Catalog #AAA9611445) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The PHD3 Antibody reacts with Human, Mouse Predicted: Pig, Bovine, Horse, Sheep, Rabbit and may cross-react with other species as described in the data sheet. AAA Biotech's PHD3 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB). WB: 1:500-1:2000. Researchers should empirically determine the suitability of the PHD3 egln3 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "PHD3, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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