EGLN3/PHD3 Antibody

Images

 
Western Blot: EGLN3/PHD3 Antibody [NB100-303] - Total protein from Mouse MEF, Human RT4 and HeLa, and Rat PC12 cells was separated on a 12% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in ...read 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 ...read more
Immunohistochemistry-Paraffin: EGLN3/PHD3 Antibody [NB100-303] - LDL Receptor was detected in immersion fixed paraffin-embedded sections of human liver cancer using rabbit anti-human antibody (Catalog # NB100-303) at ...read more
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 ...read 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 ...read more

Product Details

Summary
Reactivity Hu, Mu, Pm, RtSpecies Glossary
Applications WB, Simple Western, ChIP, IB, ICC/IF, IHC-P, IP
Clonality
Polyclonal
Host
Rabbit
Conjugate
Unconjugated
Concentration
1 mg/ml

EGLN3/PHD3 Antibody Summary

Immunogen
A synthetic peptide made to the C-terminus of humanPHD3/HIF Prolyl Hydroxylase 3. [LocusLink ID 112399]
Predicted Species
Rat (92%). Backed by our 100% Guarantee.
Isotype
IgG
Clonality
Polyclonal
Host
Rabbit
Gene
EGLN3
Purity
Immunogen affinity purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Western Blot 1:500-1:2000
  • Simple Western 1:1000
  • Chromatin Immunoprecipitation
  • Immunoblotting
  • Immunocytochemistry/Immunofluorescence
  • Immunohistochemistry-Paraffin
  • Immunoprecipitation 1:10-1:500
Application Notes
This PHD3/HIF Prolyl Hydroxylase 3 antibody is useful for Immunoprecipitation and Western blot, where a band is seen at approx. 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.Use in Immunohistochemistry-Paraffin reported in scientific literature (PMID 25161887). Use in Immunocytochemistry/immunofluorescence reported in scientific literature (PMID 20801873). Use in Immunoblotting reported in scientific literature (PMID 28366875).

In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
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.
Disclaimer note: 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.
Positive Control
HeLa Lysate (NB800-PC1)
EGLN3/PHD3 Lysate (NBL1-10156)
Reviewed Applications
Read 1 Review rated 5
using
NB100-303 in the following applications:

Publications
Read Publications using
NB100-303 in the following applications:

  • 1 publication
  • IB
    1 publication
  • 1 publication
  • 1 publication
  • IP
    1 publication
  • WB
    18 publications

Reactivity Notes

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

Packaging, Storage & Formulations

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

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
  • EGLN3
  • 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
  • pdh3
  • PHD3
  • Prolyl hydroxylase domain-containing protein 3

Background

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.

Limitations

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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NB100-303
Species: Hu, Mu, Pm, Rt
Applications: WB, Simple Western, ChIP, IB, ICC/IF, IHC-P, IP

Publications for EGLN3/PHD3 Antibody (NB100-303)(32)

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

We have publications tested in 6 applications: ChIP, IB, ICC/IF, IHC-P, IP, WB.


Filter By Application
ChIP
(1)
IB
(1)
ICC/IF
(1)
IHC-P
(1)
IP
(1)
WB
(18)
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Filter By Species
Human
(11)
Mouse
(9)
Rat
(1)
Primate
(1)
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Showing Publications 1 - 10 of 32. Show All 32 Publications.
Publications using NB100-303 Applications Species
Gibier JB, Hemon B, Fanchon M et al. Dual role of MUC1 mucin in kidney ischemia-reperfusion injury: Nephroprotector in early phase, but pro-fibrotic in late phase. Biochim. Biophys. Acta. Mar 31 2017 [PMID: 28366875] (IB) IB
Tan J, Prosser H, Dunn L et al. High-Density Lipoproteins Rescue Diabetes-Impaired Angiogenesis via Scavenger Receptor Class B Type I. Diabetes Jul 8 1905 [PMID: 27284113]
Shin J, Nunomiya A, Kitajima Y et al. Prolyl hydroxylase domain 2 deficiency promotes skeletal muscle fiber-type transition via a calcineurin/NFATc1-dependent pathway. Skelet Muscle 2016 Mar 05 [PMID: 26949511] (WB) WB
Heir P, Srikumar T, Bikopoulos G et al. Oxygen-dependent Regulation of Erythropoietin Receptor Turnover and Signaling. J Biol Chem 2016 Apr 1 [PMID: 26846855]
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)

Details:
EGLN3/PHD3 antibody used for IHC-P application on ON/10% formalin fixed visceral adipose tissues from 95 day old mice - female, male, and female ERKO/global knockout of Er alpha mice (Figure 1F).
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)

Details:
PHD3/HIF Prolyl Hydroxylase 3 antibody used for WB on human biopsy samples (vastus lateralis muscle obtained via percutaneous needle technique) in experiments involving endurance-trained /elite cyclists-triathletes and moderately active men (Figures 2B, 4 )
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
Show All 32 Publications.

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

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

Reviews using NB100-303:
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IF
(1)
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Human
(1)
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Images Ratings Applications Species Date Details
Immunofluorescence EGLN3/PHD3 NB100-303
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reviewed by:
Helder Andre
IF Human 05/14/2015
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Summary

ApplicationImmunofluorescence
Sample TestedARPE-19 cells
SpeciesHuman

Blocking

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)

Details

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 405 Labeled NB100-303AF405
Alexa Fluor 488 Labeled NB100-303AF488
Alexa Fluor 647 Labeled NB100-303AF647
Alexa Fluor 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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Recent Reviews

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Helder Andre
05/14/2015
Application: IF
Species: Human

Bioinformatics

Gene Symbol EGLN3
Entrez