Rabbit Recombinant Monoclonal HIF Prolyl Hydroxylases antibody. Carrier free. Suitable for IHC-P, IP, WB, Flow Cyt (Intra) and reacts with Human samples.
Images
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] - BSA and Azide free (AB256316), expandable thumbnail](https://content.abcam.com/products/images/hif-prolyl-hydroxylases-antibody-epr36642-25-bsa-and-azide-free-ab256316--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img54509.jpg/_jcr_content/renditions/webp-475.webp "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] - BSA and Azide free (AB256316)")
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] - BSA and Azide free (AB256316), expandable thumbnail](https://content.abcam.com/products/images/hif-prolyl-hydroxylases-antibody-epr36642-25-bsa-and-azide-free-ab256316--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img129360.jpg/_jcr_content/renditions/webp-475.webp "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] - BSA and Azide free (AB256316)")
![Immunoprecipitation - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] - BSA and Azide free (AB256316), expandable thumbnail](https://content.abcam.com/products/images/hif-prolyl-hydroxylases-antibody-epr36642-25-bsa-and-azide-free-ab256316--immunoprecipitation-img3583.jpg/_jcr_content/renditions/webp-475.webp "Immunoprecipitation - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] - BSA and Azide free (AB256316)")
![Flow Cytometry (Intracellular) - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] - BSA and Azide free (AB256316), expandable thumbnail](https://content.abcam.com/products/images/hif-prolyl-hydroxylases-antibody-epr36642-25-bsa-and-azide-free-ab256316--flow-cytometry-intracellular-img37454.jpg/_jcr_content/renditions/webp-475.webp "Flow Cytometry (Intracellular) - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] - BSA and Azide free (AB256316)")
Key facts
- Isotype
- IgG
- Host species
- Rabbit
- Storage buffer
pH: 7.2 - 7.4
Constituents: PBS- Form
- Liquid
- Clonality
- Monoclonal
Immunogen
- The exact immunogen used to generate this antibody is proprietary information.
Reactivity data
Select an application| IHC-P | IP | WB | ICC/IF | Flow Cyt (Intra) | |
|---|---|---|---|---|---|
| Human | Tested | Tested | Expected | Not recommended | Tested |
IHC-P
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info - | Notes Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
IP
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info - | Notes - |
WB
ExpectedExpected
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info Use at an assay dependent concentration. | Notes - |
ICC/IF
Not recommendedNot recommended
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info - | Notes - |
Flow Cyt (Intra)
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info - | Notes - |
Target data
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.
Alternative names
PH4, P4HTM, Transmembrane prolyl 4-hydroxylase, P4H-TM, Hypoxia-inducible factor prolyl hydroxylase 4, HIF-PH4, HIF-prolyl hydroxylase 4, HPH-4
Recommended products
Rabbit Recombinant Monoclonal HIF Prolyl Hydroxylases antibody. Carrier free. Suitable for IHC-P, IP, WB, Flow Cyt (Intra) and reacts with Human samples.
Key facts
- Isotype
- IgG
- Host species
- Rabbit
- Storage buffer
pH: 7.2 - 7.4
Constituents: PBS- Form
- Liquid
- Clonality
- Monoclonal
- Immunogen
- The exact immunogen used to generate this antibody is proprietary information.
- Carrier free
- Yes
- Clone number
- EPR3664(2)-25
- Purification technique
- Affinity purification Protein A
- Concentration
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- +4°C
- Appropriate long-term storage conditions
- +4°C
- Storage information
- Do Not Freeze
Notes
ab256316 is the carrier-free version of Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] ab255733.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
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 batch production
For more information, read more on recombinant antibodies.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
HIF prolyl hydroxylases also known as EglN or prolyl hydroxylase domain proteins (PHDs) are enzymes that modulate the stability of hypoxia-inducible factors (HIFs). These enzymes primarily facilitate the hydroxylation of prolyl residues on HIF-α subunits which marks them for degradation via the ubiquitin-proteasome pathway. Mammals express three isoforms: PHD1 PHD2 and PHD3 with PHD2 being the most widely studied. PHDs are expressed ubiquitously with higher levels observed in the liver kidneys and lung. The molecular mass of these enzymes is typically around 40 kDa.
- Biological function summary
HIF prolyl hydroxylases play an essential role in oxygen sensing and homeostasis by regulating the activity of HIF-α subunits. Under normoxic conditions the hydroxylation of HIF-α by these hydroxylases permits recognition by the von Hippel-Lindau (VHL) protein leading to proteasomal degradation. In hypoxic conditions the decreased activity of HIF prolyl hydroxylases allows HIF-α stabilization and heterodimerization with HIF-β facilitating transcription of target genes involved in angiogenesis and metabolism. PHD2 is predominantly responsible for targeting HIF-1α.
- Pathways
HIF prolyl hydroxylases integrate into the hypoxia response pathway. Oxygen-dependent regulation of HIF by these hydroxylases affects processes such as angiogenesis and erythropoiesis through interaction with several proteins including endothelial growth factor (VEGF) and erythropoietin (EPO). Beyond the hypoxia pathway they participate in metabolic pathways influencing cellular energy homeostasis thereby impacting proteins involved in metabolic regulation.
- Associated diseases and disorders
HIF prolyl hydroxylases associate with cancer and ischemic diseases. Dysregulation of these hydroxylases can lead to excessive stabilization of HIF-α which promotes tumor growth and metastasis due to enhanced angiogenesis and altered metabolism associated with cancer cells. In ischemic conditions reduced prolyl hydroxylases activity can offer protection by inducing adaptive responses through HIF-α stabilization. Their connectivity to the VHL protein further links them to von Hippel-Lindau disease characterized by tumor development in multiple organs.
Product promise
Tested
We have tested this species and application combination and it works. It is covered by our product promise.
Expected
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
Predicted
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
Not recommended
We do not recommend this combination. It is not covered by our product promise.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
4 product images
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] - BSA and Azide free (ab256316), expandable thumbnail](https://content.abcam.com/products/images/hif-prolyl-hydroxylases-antibody-epr36642-25-bsa-and-azide-free-ab256316--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img54509.jpg "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] - BSA and Azide free (ab256316)")
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] - BSA and Azide free (ab256316)
Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling HIF Prolyl Hydroxylases with Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] ab255733 at 1/250 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Cytoplasmic staining on human cerebrum (PMID: 17726031, 12163023). The section was incubated with Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] 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 (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] ab255733 ).
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] - BSA and Azide free (ab256316), expandable thumbnail](https://content.abcam.com/products/images/hif-prolyl-hydroxylases-antibody-epr36642-25-bsa-and-azide-free-ab256316--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img129360.jpg "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] - BSA and Azide free (ab256316)")
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] - BSA and Azide free (ab256316)
Immunohistochemical analysis of paraffin-embedded Human lung cancer tissue labeling HIF Prolyl Hydroxylases with Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] ab255733 at 1/250 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Cytoplasmic staining on human lung cancer (PMID: 24382781). The section was incubated with Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] 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 (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] ab255733 ).
![Immunoprecipitation - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] - BSA and Azide free (ab256316), expandable thumbnail](https://content.abcam.com/products/images/hif-prolyl-hydroxylases-antibody-epr36642-25-bsa-and-azide-free-ab256316--immunoprecipitation-img3583.jpg "Immunoprecipitation - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] - BSA and Azide free (ab256316)")
Immunoprecipitation - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] - BSA and Azide free (ab256316)
HIF Prolyl Hydroxylases was immunoprecipitated from 0.35 mg HEK-293 (human embryonic kidney epithelial cell) whole cell lysate 10μg with Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] ab255733 at 1/30 dilution. Western blot was performed on the immunoprecipitate using Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] ab255733 1/1000 dilution (0.4 μg/ml). VeriBlot for IP Detection Reagent (HRP) (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: Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] ab255733 IP in HEK-293 whole cell lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] ab255733 in HEK-293 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 1 second.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] ab255733 ).
All lanes: Immunoprecipitation - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] (Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] ab255733)
Predicted band size: 57 kDa
Observed band size: 47 kDa
![Flow Cytometry (Intracellular) - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] - BSA and Azide free (ab256316), expandable thumbnail](https://content.abcam.com/products/images/hif-prolyl-hydroxylases-antibody-epr36642-25-bsa-and-azide-free-ab256316--flow-cytometry-intracellular-img37454.jpg "Flow Cytometry (Intracellular) - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] - BSA and Azide free (ab256316)")
Flow Cytometry (Intracellular) - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] - BSA and Azide free (ab256316)
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HEK-293 (human embryonic kidney epithelial cell) cells labelling HIF Prolyl Hydroxylases with Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] ab255733 at 1/400 dilution (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] ab255733).
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