EGLN3/PHD3 Antibody


Western Blot: EGLN3/PHD3 Antibody [NB100-303] - Whole cell protein from MEF cells was separated on a 12% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed more
Immunocytochemistry/ Immunofluorescence: EGLN3/PHD3 Antibody [NB100-303] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. The cells were more
Immunocytochemistry/ Immunofluorescence: EGLN3/PHD3 Antibody [NB100-303] - analysis of PHD3 in ARPE-19 cells using anti-PHD3 antibody. Image from verified customer review.
Simple Western: EGLN3/PHD3 Antibody [NB100-303] - Simple Western lane view shows a specific band for PHD3/HIF Prolyl Hydroxylase 3 in 0.5 mg/ml of Hypoxic HeLa lysate. This experiment was performed under reducing more

Product Details

Reactivity Hu, Mu, PmSpecies Glossary
Applications WB, Simple Western, ChIP, ICC/IF, IHC-P, IP
1 mg/ml

EGLN3/PHD3 Antibody Summary

A synthetic peptide made to the C-terminus of humanPHD3/HIF Prolyl Hydroxylase 3. [LocusLink ID 112399]
Predicted Species
Rat (92%)
Immunogen affinity purified
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Packaging, Storage & Formulations

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
0.02% Sodium Azide
1 mg/ml
Immunogen affinity purified


Application Notes
This PHD3/HIF Prolyl Hydroxylase 3 antibody is useful for Immunoprecipitation and Western blot, where a band is seen at ~27 kDa. This antibody has been tested against HeLa and MEF cell lysates. Immunoprecipitation was reported in PMID: 21575608. Chromatin Immunoprecipitation was reported in scientific literature.In Simple Western only 10-15 uL of the recommended dilution is used per data point.Use in Immunohistochemistry-Paraffin reported in scientific literature (PMID 25161887). Use in Immunocytochemistry/immunofluorescence reported in scientific literature (PMID 20801873) The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Theoretical MW
27 kDa
Reviewed Applications
Read 1 Review rated 5
NB100-303 in the following application:

Read Publications using
NB100-303 in the following applications:

  • 1 publication
  • 1 publication
  • 1 publication
  • IP
    1 publication
  • WB
    17 publications

Reactivity Notes

Human and mouse. Primate reactivity reported in scientific literature (PMID: 23732909). mmunogen sequence has 92% homology to rat.

Alternate Names for EGLN3/PHD3 Antibody

  • EC 1.14.11
  • EC 1.14.11.-
  • EGL nine (C.elegans) homolog 3
  • egl nine homolog 3 (C. elegans)
  • egl nine homolog 3
  • egl nine-like protein 3 isoform
  • FLJ21620
  • HIF prolyl hydroxylase 3
  • HIFPH3 MGC125998
  • HIFPH3
  • HIF-PH3
  • HIF-prolyl hydroxylase 3
  • HPH-1
  • HPH-3
  • Hypoxia-inducible factor prolyl hydroxylase 3
  • MGC125999
  • PHD3
  • Prolyl hydroxylase domain-containing protein 3


HIF prolyl 4-hydroxylases (PHDs) are proyl hydroxylase domain-containing enzymes (PHD1/ Egln2, PHD2/ Egln1, PHD3/ Egln3, and P4H-TM) which are known for their role in mediating physiological responses to hypoxic stress via modulation of HIF1alpha expression levels. HIF-alpha subunit is regulated by hydroxylation, both by a family of PHDs leading to ubiquitination and proteasomal degradation, and by transcriptional inactivation following asparaginyl hydroxylation by FIH (factor inhibiting HIF). When oxygen levels are normal, HIF Prolyl Hydroxylase 3 (HIF-PH3/ PHD3/ Egln3) hydroxylates a specific proline found in HIF1A'a NODD/CODD domains and also hydroxylates HIF2A with preference for CODD site for HIF1A/HIF2A. Once hydroxylated, HIFs undergo proteasomal degradation via von Hippel-Lindau (VHL) ubiquitination complex. Upon hypoxic trigger, the hydroxylation reaction is tempered which let HIFs to escape degradation process leading to their nuclear translocation, heterodimerization with HIF1B, and increased expression of hypoxy-inducible genes. HIF-PH3 is the most important isozyme in limiting physiological activation of HIFs (particularly HIF2A) in hypoxia. Besides HIFs, in hypoxic conditions HIF-PH3 also hydroxylates PKM (thereby limiting glycolysis) and hydroxylates/regulates the stability of ADRB2. In cardiomyocytes, HIF-PH3 inhibits Bcl2's anti-apoptotic effect by disrupting the BAX-BCL2 complex, and in neurons, HIF-PH3 has a NGF-induced proapoptotic effect mediated via CASP3 activity regulation. HIF-PH3 also plays a crucial role in DNA damage response by hydroxylating TELO2, promoting its interaction with ATR which is required for activation of the ATR/CHK1/p53 pathway.


This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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Species: Hu, Mu, Rt, Pm
Applications: WB, Simple Western, Flow, ICC/IF, IHC, IHC-P, IP
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Applications: WB, ChIP, ELISA, Flow, GS, ICC/IF, IHC, IHC-Fr, IHC-P, IP, TCS, LA
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Applications: WB, Simple Western, EM, ICC/IF, IHC, IHC-P, IP, ICC
Species: Hu
Applications: WB, IP
Species: Hu
Applications: ICC/IF, IHC, IHC-P
Species: Hu, Mu, Rt, Fi, Ha, Pm, Sh
Applications: WB, Simple Western, ChIP, ELISA, Flow, GS, ICC/IF, IHC, IHC-Fr, IHC-P, IP
Species: Hu, Mu, Rt, Bv, Ft, Pm, Sh
Applications: WB, ChIP, GS, ICC/IF, IHC, IHC-P, IP
Species: Hu, Mu, Rt
Applications: WB, EM, ICC/IF, IHC, IHC-Fr, IHC-P, IP
Species: Hu, Mu, Rt, Bv
Applications: WB, ELISA, Flow, ICC/IF, IHC, IHC-Fr, IHC-P
Species: Hu, Mu, Rt
Applications: WB, ICC/IF, IHC, IHC-P
Species: Hu
Applications: WB
Species: Hu, Mu, Rt
Applications: WB, ICC/IF, IHC, IHC-P
Species: Hu
Applications: ICFlow
Species: Hu, Mu
Applications: WB, ELISA, ICC/IF, IHC, IHC-P
Species: Hu
Applications: WB, ICC/IF, IHC, IHC-P
Species: Hu, Mu, Pm, Rat
Applications: WB, Simple Western, ChIP, ICC/IF, IHC-P, IP

Publications for EGLN3/PHD3 Antibody (NB100-303)(28)

We have publications tested in 4 confirmed species: Human, Mouse, Rat, Primate.

We have publications tested in 5 applications: ChIP, ICC/IF, IHC-P, IP, WB.

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Showing Publications 1 - 10 of 28. Show All 28 Publications.
Publications using NB100-303 Applications Species
Zieseniss A, Hesse AR, Jatho A et al. Cardiomyocyte-Specific Transgenic Expression of Prolyl-4-Hydroxylase Domain 3 Impairs the Myocardial Response to Ischemia. Cell Physiol Biochem 2015 May 27 [PMID:26044310]
Henze At, Garvalov Bk, Seidel S et al. Loss of PHD3 allows tumours to overcome hypoxic growth inhibition and sustain proliferation through EGFR. Nat Commun. 2014 Nov 25 [PMID:25420773]
Garvalov Bk, Foss F, Henze At et al. PHD3 regulates EGFR internalization and signalling in tumours. Nat Commun. 2014 Nov 25 [PMID:25420589] (WB, Human) WB Human
Kim M, Neinast MD, Frank AP et al. ERalpha upregulates Phd3 to ameliorate HIF-1 induced fibrosis and inflammation in adipose tissue Mol Metab. 2014 Sep 01 [PMID:25161887] (IHC-P, Mouse) IHC-P Mouse
Lindholm ME, Fischer H, Poellinger L et al. Negative regulation of HIF in skeletal muscle of elite endurance athletes - a tentative mechanism promoting oxidative metabolism Am. J. Physiol. Regul. Integr. Comp. Physiol. 2014 Jun 04 [PMID:24898836] (WB, Human) WB Human
Swain Lija, Wottawa Marieke, Hillemann Annette et al. Prolyl-4-hydroxylase domain 3 (PHD3) is a critical terminator for cell survival of macrophages under stress conditions. J Leukoc Biol. 2014 Mar 13 [PMID:24626957] (Mouse) Mouse
Reischl S, Li L, Walkinshaw G et al. Inhibition of HIF prolyl-4-hydroxylases by FG-4497 reduces brain tissue injury and edema formation during ischemic stroke. PLoS ONE 2014 Jan 10 [PMID:24409307] (WB, Mouse) WB Mouse
Kim J, Kwak HJ, Cha JY et al. The role of prolyl hydroxylase domain protein (PHD) during rosiglitazone-induced adipocyte differentiation. J Biol Chem 2013 Dec 12 [PMID:24338020] (WB, Mouse) WB Mouse
Tan JT, Prosser HC, Vanags LZ et al. High-density lipoproteins augment hypoxia-induced angiogenesis via regulation of post-translational modulation of hypoxia-inducible factor 1alpha. FASEB J. 2013 Sep 10 [PMID:24022405] (WB, Human) WB Human
Fu J, Taubman MB. EGLN3 Inhibition of NF-kB Is Mediated by Prolyl Hydroxylase-independent Inhibition of IKKgamma Ubiquitination. Mol Cell Biol 2013 Jun 3 [PMID:23732909] (WB, Human, Primate) WB Human, Primate
Show All 28 Publications.

Reviews for EGLN3/PHD3 Antibody (NB100-303) (1) 51

Average Rating: 5
(Based on 1 reviews)
We have 1 review tested in 1 species: Human.

Reviews using NB100-303:
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Images Ratings Applications Species Date Details
 EGLN3/PHD3 NB100-303
reviewed by:
Human 05/14/2015


Sample TestedARPE-19 cells


Blocking Details10% goat serum in PBS-TritonX (0.1%) (30min @ RT)

Primary Anitbody

Dilution Ratio1:500 in blocking solution (1h @ RT)

Secondary Antibody

Secondary Description1:1000 Goat anti-rabbit IgG-A488 in blocking solution (1h @ RT)


Detection NotesEpifluorescence microscope
Fixation DetailsFixed in 4% buffered formalin (15min @ RT); Permeabilized in PBS-TritonX (0.5%) (10min @ RT)
Wash Description3x5min PBS

Product General Protocols

View specific protocols for EGLN3/PHD3 Antibody (NB100-303): Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol
ChIP Video Protocol
ChIP Webinar
ICC/IF Video Protocol

FAQs for EGLN3/PHD3 Antibody (NB100-303). (Showing 1 - 3 of 3 FAQs).

  1. Dear sirs, which anti-PHD3 antibody would you recommend for western blot of murine tissues? I have used the NB100-303 and I am able to detect the protein in cells, but not in heart samples. could you also recommend a protocol with Ab dilutions, solutions, transfer, etc?
    • In regards to your inquiry I am sorry to hear you have not had success with heart samples on NB100-303. We have another PHD3 antibody, NBP1-30328, that may be useful to you. It is guaranteed for mouse samples and has a sample size available if you wished to purchase and test first at a lower cost to you prior to committing to it.
  2. We are currently using the above PhD3 antibody and it is working well. However we are hoping to generate different truncation mutants of PhD3 and was wondering if the antibody will recognise these mutants. The data sheet indicates that the antibody was raised against the C-terminus of PhD3. Would it be possible to indicate where in the C-terminus the peptide is located. If the exact sequence is commercially sensitive perhaps you could tell us if the peptide is located in the fragment of 1-177 since we would like to make this mutant but are unsure if the antibody will recognise the fragment.
    • The immunogen region is located in C-terminus. Specifically the immunogen is located within the range of amino acid 177 to the C-terminus. If your truncated protein only contains amino acids 1-177, this antibody will NOT recognize this protein.
  3. Do you have any information on possible antibody dilution to use for ChIP applications?
    • We generally use around 5 x106 mammalian cells with 1 - 2 ug of NB100-303 (EGLN3/PHD3 Antibody) Therefore the amount used in ChIP with this antibody is consistent with our ChIP general protocol. 

Positive Control Lysate(s)

Secondary Antibodies


Isotype Controls

Other Available Formats

Alexa Fluor (R) 405 Labeled NB100-303AF405
Alexa Fluor (R) 488 Labeled NB100-303AF488
Alexa Fluor (R) 647 Labeled NB100-303AF647
Alexa Fluor (R) 700 Labeled NB100-303AF700
Biotin Labeled NB100-303B
DyLight 405 Labeled NB100-303V
DyLight 488 Labeled NB100-303G
DyLight 550 Labeled NB100-303R
DyLight 650 Labeled NB100-303C
FITC Labeled NB100-303F
HRP Labeled NB100-303H

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Contact Information

Product PDFs


Gene Symbol EGLN3

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