Recombinant Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] (ab255733)
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] (ab255733)](https://www.abcam.com/ps/products/255/ab255733/Images/ab255733-5-anti-hif-prolyl-hydroxylases-antibody-epr3664-25-immunohistochemistry-cerebrum-human.jpg "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] (ab255733)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR3664(2)-25] to HIF Prolyl Hydroxylases
- Suitable for: Flow Cyt (Intra), WB, IHC-P, IP
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25]
See all HIF Prolyl Hydroxylases primary antibodies -
Description
Rabbit monoclonal [EPR3664(2)-25] to HIF Prolyl Hydroxylases -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-P, IPmore details
Unsuitable for: ICC/IF -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- IHC-P: Human cerebrum and lung cancer tissue Flow Cyt (intra): HEK-293 cells IP: HEK-293 whole cell lysate WB: HEK-293 and HeLa whole cell lysates
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
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. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 0.05% BSA, 40% Glycerol (glycerin, glycerine) -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR3664(2)-25 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab255733 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 |
|---|---|---|
| Flow Cyt (Intra) |
1/400.
|
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| WB |
1/1000. Detects a band of approximately 47 kDa (predicted molecular weight: 57 kDa).
|
|
| IHC-P |
1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
|
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| IP |
1/30.
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| Notes |
|---|
|
Flow Cyt (Intra)
1/400. |
|
WB
1/1000. Detects a band of approximately 47 kDa (predicted molecular weight: 57 kDa). |
|
IHC-P
1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
|
IP
1/30. |
Target
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Function
Catalyzes the post-translational formation of 4-hydroxyproline in hypoxia-inducible factor (HIF) alpha proteins. Hydroxylates HIF1A at 'Pro-402' and 'Pro-564'. May function as a cellular oxygen sensor and, under normoxic conditions, may target HIF through the hydroxylation for proteasomal degradation via the von Hippel-Lindau ubiquitination complex. -
Tissue specificity
Widely expressed with highest levels in adult pancreas, heart, skeletal muscle, brain, placenta, kidney and adrenal gland. Expressed at lower levels in epiphyseal cartilage and in fibroblasts. -
Sequence similarities
Contains 2 EF-hand domains.
Contains 1 Fe2OG dioxygenase domain. -
Post-translational
modificationsGlycosylated. -
Cellular localization
Endoplasmic reticulum membrane. - Information by UniProt
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Database links
- Entrez Gene: 54681 Human
- SwissProt: Q9NXG6 Human
- Unigene: 654944 Human
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Alternative names
- EGLN4 antibody
- FLJ20262 antibody
- HIF prolyl hydroxylase PH4 antibody
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] (ab255733)
Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling HIF Prolyl Hydroxylases with ab255733 at 1/250 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on human cerebrum (PMID: 17726031, 12163023). The section was incubated with ab255733 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] (ab255733)](https://www.abcam.com/ps/products/255/ab255733/Images/ab255733-4-anti-hif-prolyl-hydroxylases-antibody-epr3664-25-immunohistochemistry-lung-cancer-human.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] (ab255733)Immunohistochemical analysis of paraffin-embedded Human lung cancer tissue labeling HIF Prolyl Hydroxylases with ab255733 at 1/250 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on human lung cancer (PMID: 24382781). The section was incubated with ab255733 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
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![Flow Cytometry (Intracellular) - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] (ab255733)](https://www.abcam.com/ps/products/255/ab255733/Images/ab255733-3-anti-hif-prolyl-hydroxylases-antibody-epr3664-25-flowcytometry-hek293-human.jpg)
Flow Cytometry (Intracellular) - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] (ab255733)Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HEK-293 (human embryonic kidney epithelial cell) cells labelling HIF Prolyl Hydroxylases with ab255733 at 1/400 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).
Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
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![Immunoprecipitation - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] (ab255733)](https://www.abcam.com/ps/products/255/ab255733/Images/ab255733-2-anti-hif-prolyl-hydroxylases-antibody-epr3664-25-immunoprecipitation-hek293-human.jpg)
Immunoprecipitation - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] (ab255733)HIF Prolyl Hydroxylases was immunoprecipitated from 0.35 mg HEK-293 (human embryonic kidney epithelial cell) whole cell lysate 10μg with ab255733 at 1/30 dilution. Western blot was performed on the immunoprecipitate using ab255733 1/1000 dilution (0.4 μg/ml). VeriBlot for IP Detection Reagent (HRP) (ab131366) was used as the secondary antibody at 1/5000 dilution.
Lane 1: HEK-293 (human embryonic kidney epithelial cell) whole cell lysate 10μg
Lane 2: ab255733 IP in HEK-293 whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab255733 in HEK-293 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 1 second.
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![Western blot - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] (ab255733)](https://www.abcam.com/ps/products/255/ab255733/Images/ab255733-1-anti-hif-prolyl-hydroxylases-antibody-epr3664-25-western-bot-human.jpg)
Western blot - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] (ab255733)All lanes : Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] (ab255733) at 1/1000 dilution
Lane 1 : HEK-293 (human embryonic kidney epithelial cell) whole cell lysate
Lane 2 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 57 kDa
Observed band size: 47 kDa why is the actual band size different from the predicted?
Exposure time: 20 secondsBlocking/Diluting buffer and concentration: 5% NFDM/TBST.
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![Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] (ab255733)](https://www.abcam.com/ps/products/255/ab255733/Images/ab255733-6-benefits-of-recombinant-antibodies.png)
Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] (ab255733)
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (0)
ab255733 has not yet been referenced specifically in any publications.