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AB256316

Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] - BSA and Azide free

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Rabbit Recombinant Monoclonal HIF Prolyl Hydroxylases antibody. Carrier free. Suitable for IHC-P, IP, WB, Flow Cyt (Intra) and reacts with Human samples.

View Alternative Names

PH4, P4HTM, Transmembrane prolyl 4-hydroxylase, P4H-TM, Hypoxia-inducible factor prolyl hydroxylase 4, HIF-PH4, HIF-prolyl hydroxylase 4, HPH-4

4 Images
Flow Cytometry (Intracellular) - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] - BSA and Azide free (AB256316)
  • Flow Cyt (Intra)

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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 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab255733).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] - BSA and Azide free (AB256316)
  • IHC-P

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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 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).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab255733 ).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] - BSA and Azide free (AB256316)
  • IHC-P

Unknown

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 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).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab255733 ).

Immunoprecipitation - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] - BSA and Azide free (AB256316)
  • IP

Unknown

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 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab255733 ).

All lanes:

Immunoprecipitation - Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] (<a href='/en-us/products/primary-antibodies/hif-prolyl-hydroxylases-antibody-epr36642-25-ab255733'>ab255733</a>)

Predicted band size: 57 kDa

Observed band size: 47 kDa

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  • Unconjugated

    Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25]

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR3664(2)-25

Isotype

IgG

Carrier free

Yes

Reacts with

Human

Applications

IP, Flow Cyt (Intra), WB, IHC-P

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

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Product details

ab256316 is the carrier-free version of ab255733.

Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

What are the advantages of a recombinant monoclonal antibody?
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.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. 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.

Compatibility
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.

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Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.2 - 7.4 Constituents: PBS
Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
Do Not Freeze
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

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.

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.
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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:
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Target data

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.
See full target information P4HTM
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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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