antibody

Anti-PHD3/EGLN3 Picoband Antibody

A02182-1

100 ug/vial

polyclonal

domestic rabbit

nonconjugated

human, mouse, rat

western blot, immunohistochemistry

Enabled

Anti-PHD3/EGLN3 Picoband Antibody

Primary Antibodies, Polyclonal Antibodies, IHC ICC IF Antibodies

EGLN3

Unconjugated / $280 APC / $330 APC-Cy7 / $330 FITC / $330 PE / $330 PE-Cy5 / $330 PE-Cy7 / $330 100ug / $280 100ug+Free HRP Secondary BA1054 / $280 100ug+Free Biotin Secondary BA1003 / $280

Polyclonal

0.5-1mg/ml, actual concentration vary by lot. Use suggested dilution ratio to decide dilution procedure.

No

Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na 2 HPO 4 , 0.05mg NaN 3 .

Rabbit IgG polyclonal antibody for PHD3 detection. Tested with WB, IHC-P in Human;Mouse;Rat.

Rabbit IgG polyclonal antibody for PHD3 detection. Tested with WB, IHC-P in Human;Mouse;Rat.

100 ug/vial

Q9H6Z9

Rabbit

A synthetic peptide corresponding to a sequence of human PHD3 (ATRYAMTVWYFDAEERAEAKKKFRNLTRKTESALTED).

Lyophilized

At -20 C for one year. After reconstitution, at 4 C for one month. It can also be aliquotted and stored frozen at -20 C for a longer time. Avoid repeated freezing and thawing.

No cross reactivity with other proteins.

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

Western blot, 0.1-0.5ug/ml. Immunohistochemistry(Paraffin-embedded Section), 0.5-1ug/ml.

IHC, WB

Human, Mouse, Rat

Figure 1. Western blot analysis of PHD3 using anti-PHD3 antibody (A02182-1). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions. Lane 1: human placenta tissue lysates, . Lane 2: mouse lung tissue lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-PHD3 antigen affinity purified polyclonal antibody (Catalog # A02182-1) at 0.5 ?g/mL overnight at 4?C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for PHD3 at approximately 27KD. The expected band size for PHD3 is at 27KD. Figure 2. IHC analysis of PHD3 using anti-PHD3 antibody (A02182-1). PHD3 was detected in paraffin-embedded section of human Lung cancer tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2?g/ml rabbit anti-PHD3 Antibody (A02182-1) overnight at 4?C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37?C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen. Figure 3. IHC analysis of PHD3 using anti-PHD3 antibody (A02182-1). PHD3 was detected in paraffin-embedded section of human tonsil tissue . Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2?g/ml rabbit anti-PHD3 Antibody (A02182-1) overnight at 4?C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37?C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen. Figure 4. IHC analysis of PHD3 using anti-PHD3 antibody (A02182-1). PHD3 was detected in paraffin-embedded section of mouse lung tissue . Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-PHD3 Antibody (A02182-1) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen. Figure 5. IHC analysis of PHD3 using anti-PHD3 antibody (A02182-1). PHD3 was detected in paraffin-embedded section of rat skeletal muscle tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-PHD3 Antibody (A02182-1) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

Egl nine homolog 3, also known as PHD3, is a protein that in humans is encoded by the EGLN3 gene. ELGN3 is a member of the superfamily of non-haem Fe(II) and 2-oxoglutarate (2OG)-dependent dioxygenases. It is mapped to chromosome 14q13.1 based on an alignment of the EGLN3 sequence with the genomic sequence (GRCh37). ELGN3 is a widely expressed 27 kDa enzyme that hydroxylates proline residues on target proteins including HIF-1 alpha.

Invasion/Microenvironment, Metabolism, Metabolism Processes, Pathways And Processes, Response To Hypoxia

Egl nine homolog 3; HPH-1; Hypoxia-inducible factor prolyl hydroxylase 3; HIF-PH3; HIF-prolyl hydroxylase 3; HPH-3; Prolyl hydroxylase domain-containing protein 3; PHD3; EGLN3

egl-9 family hypoxia inducible factor 3

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.

Nucleus. Cytoplasm. Colocalizes with WDR83 in the cytoplasm.

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.

Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit (SV0002-1) for IHC(P).

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