Recombinant Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] (ab133630)
- Datasheet
- References (1)
- Protocols
Overview
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Product name
Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)]
See all PHD2 / prolyl hydroxylase primary antibodies -
Description
Rabbit monoclonal [EPR3660(B)(2)] to PHD2 / prolyl hydroxylase -
Host species
Rabbit -
Tested applications
Suitable for: WB, IPmore details
Unsuitable for: ICC or IHC-P -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human PHD2/ prolyl hydroxylase (N terminal). The exact sequence is proprietary.
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Positive control
- SH-SY5Y treated with cobalt chloride, SH-SY5Y, and Human adrenal gland lysates
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General notes
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
This product is a recombinant rabbit monoclonal antibody.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Dissociation constant (KD)
KD = 1.43 x 10 -11 M Learn more about KD -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR3660(B)(2) -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab133630 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 |
|---|---|---|
| WB | 1/1000 - 1/10000. Predicted molecular weight: 46 kDa. | |
| IP | 1/10 - 1/100. |
Target
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Function
Catalyzes the post-translational formation of 4-hydroxyproline in hypoxia-inducible factor (HIF) alpha proteins. Hydroxylates HIF-1 alpha at 'Pro-402' and '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. -
Tissue specificity
According to PubMed:11056053, widely expressed with highest levels in skeletal muscle and heart, moderate levels in pancreas, brain (dopaminergic neurons of adult and fetal substantia nigra) and kidney, and lower levels in lung and liver. According to PubMed:12351678 widely expressed with highest levels in brain, kidney and adrenal gland. Expressed in cardiac myocytes, aortic endothelial cells and coronary artery smooth muscle. -
Involvement in disease
Defects in EGLN1 are the cause of erythrocytosis familial type 3 (ECYT3) [MIM:609820]. ECYT3 is an autosomal dominant disorder characterized by increased serum red blood cell mass, elevated serum hemoglobin and hematocrit, and normal serum erythropoietin levels. -
Sequence similarities
Contains 1 Fe2OG dioxygenase domain.
Contains 1 MYND-type zinc finger. - Information by UniProt
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Database links
- Entrez Gene: 54583 Human
- Omim: 606425 Human
- SwissProt: Q9GZT9 Human
- Unigene: 444450 Human
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Alternative names
- C1ORF12 antibody
- Chromosome 1 Open Reading Frame 12 antibody
- DKFZp761F179 antibody
see all
Images
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![Western blot - Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] (ab133630)](https://www.abcam.com/ps/products/133/ab133630/Images/ab133630-258296-anti-phd2-prolyl-hydroxylase-antibody-epr3660b2-western-blot.jpg)
Western blot - Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] (ab133630)Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: PHD2 / prolyl hydroxylase knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: HepG2 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab133630 observed at 1X kDa. Red - loading control, ab8245, observed at 37 kDa.
Unpurified ab133630 was shown to specifically react with PHD2 / prolyl hydroxylase when PHD2 / prolyl hydroxylase knockout samples were used. Wild-type and PHD2 / prolyl hydroxylase knockout samples were subjected to SDS-PAGE. Unpurified ab133630 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging. -
![Immunoprecipitation - Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] (ab133630)](https://www.abcam.com/ps/products/133/ab133630/Images/ab133630-310272-anti-phd2-prolyl-hydroxylase-antibody-epr3660b2-immunoprecipitation.jpg)
Immunoprecipitation - Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] (ab133630)ab133630 (purified) at 1:40 dilution (2µg) immunoprecipitating PHD2 / prolyl hydroxylase in SH-SY5Y treated with 0.1mM cobalt chloride for 8 hours whole cell lysate.
Lane 1 (input): SH-SY5Y (Human neuroblastoma epithelial cell) treated with 0.1mM cobalt chloride for 8 hours whole cell lysate 10µg
Lane 2 (+): ab133630 & SH-SY5Y treated with 0.1mM cobalt chloride for 8 hours whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab133630 in SH-SY5Y treated with 0.1mM cobalt chloride for 8 hours whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST. -
![Other - Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] (ab133630)](https://www.abcam.com/ps/products/133/ab133630/Images/ab133630-147524-anti-phd2-prolyl-hydroxylase-antibody-epr3660b2.jpg)
Other - Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] (ab133630) -
![Western blot - Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] (ab133630)](https://www.abcam.com/ps/products/133/ab133630/Images/ab133630-310240-anti-phd2-prolyl-hydroxylase-antibody-epr3660b2-western-blot.jpg)
Western blot - Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] (ab133630)All lanes : Anti-PHD2 / prolyl hydroxylase antibody [EPR3660(B)(2)] (ab133630) at 1/5000 dilution (Purified)
Lane 1 : SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysates with 5% NFDM/TBST
Lane 2 : SH-SY5Y (Human neuroblastoma epithelial cell) treated with 0.1mM cobalt chloride for 8 hours whole cell lysates with 5% NFDM/TBST
Lane 3 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates with 5% NFDM/TBST
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 46 kDa
Observed band size: 46 kDa
Datasheets and documents
References
This product has been referenced in:
- Bartoszewska S et al. The hypoxia-inducible miR-429 regulates hypoxia-inducible factor-1a expression in human endothelial cells through a negative feedback loop. FASEB J 29:1467-79 (2015). Read more (PubMed: 25550463) »
