AB184714

Anti-PHD3 antibody [EPR17869]

RabMAb
Recombinant
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(11 Publications)

Rabbit Recombinant Monoclonal PHD3 antibody. Suitable for IP, WB, ICC/IF and reacts with Mouse, Rat, Human samples. Cited in 11 publications.

View Alternative Names

Prolyl hydroxylase EGLN3, Egl nine homolog 3, HPH-1, Hypoxia-inducible factor prolyl hydroxylase 3, Prolyl hydroxylase domain-containing protein 3, HIF-PH3, HIF-prolyl hydroxylase 3, HPH-3, PHD3, EGLN3

9 Images

Immunocytochemistry/ Immunofluorescence - Anti-PHD3 antibody [EPR17869] (AB184714)

ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-PHD3 antibody [EPR17869] (AB184714)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A549 (Human lung carcinoma) cells labeling PHD3 with ab184714 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing weakly cytoplasm and nuclear staining on A549 cell line.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

The negative controls are as follows : -

-ve control 1 : ab184714 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.

-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-PHD3 antibody [EPR17869] (AB184714)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized PC-12 (Rat adrenal gland pheochromocytoma) cells labeling PHD3 with ab184714 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing weakly cytoplasm and nuclear staining on PC-12 cells.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

The negative controls are as follows : -

-ve control 1 : ab184714 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.

-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

Supplier Data

Immunoprecipitation - Anti-PHD3 antibody [EPR17869] (AB184714)

PHD3 was immunoprecipitated from 1mg of NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate with ab184714 at 1/70 dilution.

Western blot was performed from the immunoprecipitate using ab184714 at 1/1000 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/10000 dilution.

Lane 1 : NIH/3T3 whole cell lysate 10ug (Input).

Lane 2 : ab184714 IP in NIH/3T3 whole cell lysate.

Lane 3 : NIH/3T3 whole cell lysate supernatant after capture (unbound).

Lane 4 : Rabbit monoclonal IgG (ab172730) instead of ab184714 in NIH/3T3 whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 30 seconds.

ab184714 is not a strong binder for IP - only a partial amount of the target protein in the lysate was immune-precipitated.

All lanes:

Immunoprecipitation - Anti-PHD3 antibody [EPR17869] (ab184714)

Predicted band size: 27 kDa

false

Supplier Data

Western blot - Anti-PHD3 antibody [EPR17869] (AB184714)

Blocking/Dilution buffer : 5% NFDM/TBST.

PHD3 expression was induced by CoCl2 treatment (PMID : 18337469).

All lanes:

Western blot - Anti-PHD3 antibody [EPR17869] (ab184714) at 1/2000 dilution

Lane 1:

Untreated MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 10 µg

Lane 2:

MCF7 cell lysate treated with 0.5mM CoCl2 (Cobalt (II) chloride) for 6 hours at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 27 kDa

Observed band size: 27 kDa

false

Exposure time: 1min

Supplier Data

Western blot - Anti-PHD3 antibody [EPR17869] (AB184714)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-PHD3 antibody [EPR17869] (ab184714) at 1/2000 dilution

All lanes:

Human fetal liver lysate at 10 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 27 kDa

Observed band size: 27 kDa

false

Exposure time: 15s

Supplier Data

Western blot - Anti-PHD3 antibody [EPR17869] (AB184714)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-PHD3 antibody [EPR17869] (ab184714) at 1/5000 dilution

All lanes:

A549 (Human lung carcinoma) whole cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 27 kDa

Observed band size: 27 kDa

false

Exposure time: 2min

Supplier Data

Western blot - Anti-PHD3 antibody [EPR17869] (AB184714)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-PHD3 antibody [EPR17869] (ab184714) at 1/2000 dilution

Lane 1:

Mouse pancreas lysate at 10 µg

Lane 2:

Rat pancreas lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 27 kDa

Observed band size: 27 kDa

false

Exposure time: 1min

Supplier Data

Western blot - Anti-PHD3 antibody [EPR17869] (AB184714)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-PHD3 antibody [EPR17869] (ab184714) at 1/2000 dilution

Lane 1:

Mouse kidney lysate at 10 µg

Lane 2:

Mouse spleen lysate at 10 µg

Lane 3:

Rat brain lysate at 10 µg

Lane 4:

RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg

Lane 5:

PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate at 10 µg

Lane 6:

NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 27 kDa

Observed band size: 27 kDa

false

Exposure time: 15s

Supplier Data

Western blot - Anti-PHD3 antibody [EPR17869] (AB184714)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-PHD3 antibody [EPR17869] (ab184714) at 1/10000 dilution

Lane 1:

PHD3 transfected HEK-293 (Human epithelial cells from embryonic kidney) whole cell lysate at 10 µg

Lane 2:

Empty vector (vector control) transfected HEK-293 whole cell lysate at 10 µg

Lane 3:

PHD3 transfected HEK-293 whole cell lysate treated with 0.1 mM CoCl2 for 4 hours at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 27 kDa

Observed band size: 27 kDa

false

Exposure time: 1s

665 Alexa Fluor® 647

Alexa Fluor® 647 Anti-PHD3 antibody [EPR17869]
Carrier free

Anti-PHD3 antibody [EPR17869] - BSA and Azide free

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR17869

Isotype

IgG

Carrier free

Reacts with

Mouse, Rat, Human

Applications

WB, ICC/IF, IP

applications

Immunogen

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

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "IP-species-checked": "guaranteed", "IP-species-dilution-info": "", "IP-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/2000", "WB-species-notes": "", "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "1/250", "ICCIF-species-notes": "" }, "Mouse": { "IP-species-checked": "testedAndGuaranteed", "IP-species-dilution-info": "1/70", "IP-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/2000", "WB-species-notes": "", "ICCIF-species-checked": "guaranteed", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "" }, "Rat": { "IP-species-checked": "guaranteed", "IP-species-dilution-info": "", "IP-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/2000", "WB-species-notes": "", "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "1/250", "ICCIF-species-notes": "" } } }

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Product details

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.

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Properties and storage information

Form

Liquid

Purification technique

Affinity purification Protein A

Storage buffer

pH: 7.2 Preservative: 0.01% Sodium azide Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

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Supplementary information

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

The PHD3 protein also known as EGLN3 or Prolyl Hydroxylase Domain-Containing Protein 3 functions mechanically to regulate oxygen homeostasis in cells. It catalyzes the hydroxylation of proline residues on hypoxia-inducible transcription factors (HIFs). The molecular mass of PHD3 is approximately 27 kDa. PHD3 expresses in a variety of tissues notably in the heart brain and skeletal muscles. Its expression often occurs in response to hypoxic conditions reflecting its role in oxygen sensing and adaptation to change in oxygen levels.

Biological function summary

The PHD3 protein plays an essential role in regulating the degradation of HIFs preventing their accumulation under normoxic conditions. It is part of a larger complex which includes oxygen iron and 2-oxoglutarate facilitating its hydroxylase activity. Hydroxylation of HIFs by PHD3 marks them for degradation via the ubiquitin-proteasome pathway preventing HIFs from activating genes related to erythropoiesis angiogenesis and cellular metabolism adaptation to hypoxia. Through these actions PHD3 helps maintain cellular oxygen homeostasis and metabolic balance.

Pathways

PHD3 is integral to the HIF signaling pathway and the cellular response to hypoxia. Its interaction with HIF-1α and HIF-2α is important in this context dictating the stability and activity of these transcription factors under varying oxygen levels. PHD3 also associates with other prolyl hydroxylases such as PHD1 and PHD2 coordinating the regulation of HIFs collectively across different cell types and conditions. These interactions contribute to the modulation of gene expression in response to hypoxic stress.

Aberrant PHD3 activity links to cancer and ischemic diseases. In cancer altered PHD3 expression affects tumor growth and metastasis by disrupting normal oxygen sensing allowing cancer cells to adapt to low-oxygen environments. Moreover PHD3's interaction with proteins like HIF-1α and HIF-2α plays a role in the pathological angiogenesis seen in certain cancer types. In ischemic diseases improper regulation by PHD3 might impede normal tissue responses to reduced blood flow affecting recovery. Its specific modulation in diseases presents potential therapeutic targets for drug development.

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Product protocols

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

Visit the General protocols
Visit the Troubleshooting

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Target data

Prolyl hydroxylase that mediates hydroxylation of proline residues in target proteins, such as PKM, TELO2, ATF4 and HIF1A (PubMed : 19584355, PubMed : 20978507, PubMed : 21483450, PubMed : 21575608, PubMed : 21620138, PubMed : 22797300). Target proteins are preferentially recognized via a LXXLAP motif. Cellular oxygen sensor that catalyzes, under normoxic conditions, the post-translational formation of 4-hydroxyproline in hypoxia-inducible factor (HIF) alpha proteins (PubMed : 11595184, PubMed : 12181324). Hydroxylates a specific proline found in each of the oxygen-dependent degradation (ODD) domains (N-terminal, NODD, and C-terminal, CODD) of HIF1A (PubMed : 11595184, PubMed : 12181324). Also hydroxylates HIF2A (PubMed : 11595184, PubMed : 12181324). Has a preference for the CODD site for both HIF1A and HIF2A (PubMed : 11595184, PubMed : 12181324). Hydroxylation on the NODD site by EGLN3 appears to require prior hydroxylation on the CODD site (PubMed : 11595184, PubMed : 12181324). Hydroxylated HIFs are then targeted for proteasomal degradation via the von Hippel-Lindau ubiquitination complex (PubMed : 11595184, PubMed : 12181324). Under hypoxic conditions, the hydroxylation reaction is attenuated allowing HIFs to escape degradation resulting in their translocation to the nucleus, heterodimerization with HIF1B, and increased expression of hypoxy-inducible genes (PubMed : 11595184, PubMed : 12181324). ELGN3 is the most important isozyme in limiting physiological activation of HIFs (particularly HIF2A) in hypoxia. Also hydroxylates PKM in hypoxia, limiting glycolysis (PubMed : 21483450, PubMed : 21620138). Under normoxia, hydroxylates and regulates the stability of ADRB2 (PubMed : 19584355). Regulator of cardiomyocyte and neuronal apoptosis. In cardiomyocytes, inhibits the anti-apoptotic effect of BCL2 by disrupting the BAX-BCL2 complex (PubMed : 20849813). In neurons, has a NGF-induced proapoptotic effect, probably through regulating CASP3 activity (PubMed : 16098468). Also essential for hypoxic regulation of neutrophilic inflammation (PubMed : 21317538). Plays a crucial role in DNA damage response (DDR) by hydroxylating TELO2, promoting its interaction with ATR which is required for activation of the ATR/CHK1/p53 pathway (PubMed : 22797300). Also mediates hydroxylation of ATF4, leading to decreased protein stability of ATF4 (Probable).

See full target information EGLN3

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Publications (11)

Recent publications for all applications. Explore the full list and refine your search

Molecular medicine (Cambridge, Mass.) 31:100 PubMed40087582

2025

Sirtuin1 mitigates hypoxia-induced cardiomyocyte apoptosis in myocardial infarction via PHD3/HIF-1α.

Applications

Unspecified application

Species

Unspecified reactive species

Yafen Chen,Shuyao Shan,Qiqi Xue,Yan Ren,Qihong Wu,Jiawei Chen,Ke Yang,Jiumei Cao

Respiratory research 26:61 PubMed39985019

2025

Mechanistic insights into the role of EGLN3 in pulmonary vascular remodeling and endothelial dysfunction.

Applications

Unspecified application

Species

Unspecified reactive species

Xiaodong Deng,Qing Que,Kunchi Zhang,Bo Li,Nianlong Yang,Qiang Hu,Sheng Lv,Yi Liu

Frontiers in physiology 15:1462014 PubMed39469441

2024

Unveiling the proteome of the fasting heart: Insights into HIF-1 pathway regulation.

Applications

Unspecified application

Species

Unspecified reactive species

Daniel Benak,Kristyna Holzerova,Frantisek Kolar,Miloslava Chalupova,Marketa Hlavackova

Journal of nanobiotechnology 22:479 PubMed39134988

2024

Oral administration of Robinia pseudoacacia L. flower exosome-like nanoparticles attenuates gastric and small intestinal mucosal ferroptosis caused by hypoxia through inhibiting HIF-1α- and HIF-2α-mediated lipid peroxidation.

Applications

Unspecified application

Species

Unspecified reactive species

Dezhi Wang,Heng Zhang,Xingchen Liao,Jun Li,Jie Zeng,Yilin Wang,Mingjie Zhang,Xianzong Ma,Xin Wang,Fangli Ren,Yinyin Wang,Meng Li,Junfeng Xu,Peng Jin,Jianqiu Sheng

Heliyon 10:e33206 PubMed39021988

2024

Comprehensive analysis and experimental verification reveal the molecular characteristics of EGLN3 in pan-cancer and its relationship with the proliferation and apoptosis of lung cancer.

Applications

Unspecified application

Species

Unspecified reactive species

Yuan-Xiang Shi,Peng-Hui Dai,Tao Chen,Jian-Hua Yan

Journal of translational medicine 22:248 PubMed38454480

2024

Inhibition of OGFOD1 by FG4592 confers neuroprotection by activating unfolded protein response and autophagy after ischemic stroke.

Applications

Unspecified application

Species

Unspecified reactive species

Jian Xie,Yuan Zhang,Bin Li,Wen Xi,Yu Wang,Lu Li,Chenchen Liu,Ling Shen,Bing Han,Yan Kong,HongHong Yao,Zhijun Zhang

The EMBO journal 43:931-955 PubMed38360997

2024

VHL suppresses autophagy and tumor growth through PHD1-dependent Beclin1 hydroxylation.

Applications

Unspecified application

Species

Unspecified reactive species

Zheng Wang,Meisi Yan,Leiguang Ye,Qimin Zhou,Yuran Duan,Hongfei Jiang,Lei Wang,Yuan Ouyang,Huahe Zhang,Yuli Shen,Guimei Ji,Xiaohan Chen,Qi Tian,Liwei Xiao,Qingang Wu,Ying Meng,Guijun Liu,Leina Ma,Bo Lei,Zhimin Lu,Daqian Xu

The British journal of dermatology 187:936-947 PubMed35862273

2022

Insights into male androgenetic alopecia using comparative transcriptome profiling: hypoxia-inducible factor-1 and Wnt/β-catenin signalling pathways.

Applications

Unspecified application

Species

Unspecified reactive species

Qingmei Liu,Yulong Tang,Yan Huang,Ji'an Wang,Kai Yang,Yuting Zhang,Weilin Pu,Jing Liu,Xiangguang Shi,Yanyun Ma,Chunya Ni,Yue Zhang,Yifei Zhu,Haiyang Li,Jiucun Wang,Jinran Lin,Wenyu Wu

International journal of molecular sciences 22: PubMed33799686

2021

Prolyl Hydroxylase 3 Knockdown Accelerates -Mutant Kidney Cancer Growth In Vivo.

Applications

Unspecified application

Species

Unspecified reactive species

Niki M Zacharias,Lei Wang,Tapati Maity,Li Li,Steven W Millward,Jose A Karam,Christopher G Wood,Neema Navai

The Journal of biological chemistry 295:16299-16313 PubMed32963106

2020

TET is targeted for proteasomal degradation by the PHD-pVHL pathway to reduce DNA hydroxymethylation.

Applications

Unspecified application

Species

Unspecified reactive species

Sijia Fan,Jing Wang,Guangqing Yu,Fangjing Rong,Dawei Zhang,Chenxi Xu,Juan Du,Zhi Li,Gang Ouyang,Wuhan Xiao

View all publications

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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'.

For licensing inquiries, please contact partnerships@abcam.com

Anti-PHD3 antibody [EPR17869]

Immunocytochemistry/ Immunofluorescence - Anti-PHD3 antibody [EPR17869] (AB184714)

Immunocytochemistry/ Immunofluorescence - Anti-PHD3 antibody [EPR17869] (AB184714)

Immunoprecipitation - Anti-PHD3 antibody [EPR17869] (AB184714)

Western blot - Anti-PHD3 antibody [EPR17869] (AB184714)

Western blot - Anti-PHD3 antibody [EPR17869] (AB184714)

Western blot - Anti-PHD3 antibody [EPR17869] (AB184714)

Western blot - Anti-PHD3 antibody [EPR17869] (AB184714)

Western blot - Anti-PHD3 antibody [EPR17869] (AB184714)

Western blot - Anti-PHD3 antibody [EPR17869] (AB184714)

Related conjugates and formulations (2)

Key facts

Host species

Clonality

Clone number

Isotype

Carrier free

Reacts with

Applications

Immunogen

Reactivity data

Product details

Properties and storage information

Form

Purification technique

Storage buffer

Shipped at conditions

Appropriate short-term storage duration

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Aliquoting information

Storage information

Supplementary information

Biological function summary

Pathways

Product protocols

Target data

Publications (11)

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