Anti-PHD3 antibody (ab30782)
Key features and details
- Rabbit polyclonal to PHD3
- Suitable for: ICC/IF, WB
- Reacts with: Human
- Isotype: IgG
Overview
-
Product name
Anti-PHD3 antibody
See all PHD3 primary antibodies -
Description
Rabbit polyclonal to PHD3 -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, WBmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat
-
Immunogen
Synthetic peptide corresponding to Human PHD3 aa 50-150 (C terminal) conjugated to keyhole limpet haemocyanin.
(Peptide available asab30781)
Properties
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
-
Compatible Secondaries
-
Isotype control
-
Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab30782 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application Abreviews Notes ICC/IF Use a concentration of 5 µg/ml. WB Use a concentration of 1 µg/ml. Detects a band of approximately 27 kDa (predicted molecular weight: 27 kDa). Target
-
Function
Catalyzes the post-translational formation of 4-hydroxyproline in hypoxia-inducible factor (HIF) alpha proteins. Hydroxylates HIF-1 alpha at 'Pro-564', and HIF-2 alpha. Functions as a cellular oxygen sensor and, under normoxic conditions, targets HIF through the hydroxylation for proteasomal degradation via the von Hippel-Lindau ubiquitination complex. May play a role in cell growth regulation in muscle cells and in apoptosis in neuronal tissue. Promotes cell death through a caspase-dependent mechanism. -
Tissue specificity
Widely expressed at low levels. Expressed at higher levels in heart (cardiac myocytes, aortic endothelial cells and coronary artery smooth muscle) and placenta. -
Sequence similarities
Contains 1 Fe2OG dioxygenase domain. -
Cellular localization
Cytoplasm. Nucleus. - Information by UniProt
-
Database links
- Entrez Gene: 112399 Human
- Entrez Gene: 112407 Mouse
- Entrez Gene: 54702 Rat
- Omim: 606426 Human
- SwissProt: Q9H6Z9 Human
- SwissProt: Q91UZ4 Mouse
- SwissProt: Q62630 Rat
- Unigene: 135507 Human
see all -
Alternative names
- Egl 9 family hypoxia inducible factor 3 antibody
- Egl nine homolog 3 (C. elegans) antibody
- Egl nine homolog 3 antibody
see all
Images
-
Western blot - Anti-PHD3 antibody (ab30782)All lanes : Anti-PHD3 antibody (ab30782) at 1 µg/ml
Lane 1 :Hela-Vehicle treated (Negative Control) Whole Cell Lysate (ab116321)
Lane 2 : Hela-DFO treated (0.5mM, 24h) Whole Cell Lysate (ab116322)
Lane 3 : Human HeLa Nuclear DFO treated
Lane 4 : Human HeLa Cytoplasmic DFO treated
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 27 kDa
Observed band size: 27 kDa
Additional bands at: 18 kDa (possible non-specific binding)
Exposure time: 20 minutesThis blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab30782 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.
-
Western blot - Anti-PHD3 antibody (ab30782)This image is courtesy of an anonymous AbreviewAll lanes : Anti-PHD3 antibody (ab30782) at 1/1000 dilution
Lane 1 : Mouse embroyonic fibroblast cell lysate. Treated 21% O2 for 24 hours
Lane 2 : Mouse embroyonic fibroblast cell lysate. Treated 1% O2 for 24 hours
Lysates/proteins at 5 µg per lane.
Secondary
All lanes : Goat polyclonal anti-rabbit HRP-conjugate at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 27 kDa
Exposure time: 30 seconds
-
Immunocytochemistry/ Immunofluorescence - Anti-PHD3 antibody (ab30782)ICC/IF image of ab30782 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab30782 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Protocols
Datasheets and documents
References (12)
ab30782 has been referenced in 12 publications.
- Tsai YM et al. Loss of miR-145-5p Causes Ceruloplasmin Interference with PHD-Iron Axis and HIF-2a Stabilization in Lung Adenocarcinoma-Mediated Angiogenesis. Int J Mol Sci 21:N/A (2020). PubMed: 32708433
- Lee YM et al. Thymoquinone Selectively Kills Hypoxic Renal Cancer Cells by Suppressing HIF-1a-Mediated Glycolysis. Int J Mol Sci 20:N/A (2019). PubMed: 30832444
- Li D et al. Armadillo repeat containing 12 promotes neuroblastoma progression through interaction with retinoblastoma binding protein 4. Nat Commun 9:2829 (2018). PubMed: 30026490
- Tang LR et al. Prolyl hydroxylase domain 3 influences the radiotherapy efficacy of pancreatic cancer cells by targeting hypoxia-inducible factor-1a. Onco Targets Ther 11:8507-8515 (2018). PubMed: 30555241
- Xia YJ et al. PHD3 affects gastric cancer progression by negatively regulating HIF1A. Mol Med Rep 16:6882-6889 (2017). PubMed: 28901473
Images
-
Western blot - Anti-PHD3 antibody (ab30782)All lanes : Anti-PHD3 antibody (ab30782) at 1 µg/ml
Lane 1 :Hela-Vehicle treated (Negative Control) Whole Cell Lysate (ab116321)
Lane 2 : Hela-DFO treated (0.5mM, 24h) Whole Cell Lysate (ab116322)
Lane 3 : Human HeLa Nuclear DFO treated
Lane 4 : Human HeLa Cytoplasmic DFO treated
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 27 kDa
Observed band size: 27 kDa
Additional bands at: 18 kDa (possible non-specific binding)
Exposure time: 20 minutesThis blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab30782 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.
-
Western blot - Anti-PHD3 antibody (ab30782) This image is courtesy of an anonymous AbreviewAll lanes : Anti-PHD3 antibody (ab30782) at 1/1000 dilution
Lane 1 : Mouse embroyonic fibroblast cell lysate. Treated 21% O2 for 24 hours
Lane 2 : Mouse embroyonic fibroblast cell lysate. Treated 1% O2 for 24 hours
Lysates/proteins at 5 µg per lane.
Secondary
All lanes : Goat polyclonal anti-rabbit HRP-conjugate at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 27 kDa
Exposure time: 30 seconds
-
Immunocytochemistry/ Immunofluorescence - Anti-PHD3 antibody (ab30782)
ICC/IF image of ab30782 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab30782 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
