EGLN3/PHD3 Antibody

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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
Simple Western: EGLN3/PHD3 Antibody [NB100-303] - Image shows a specific band for PHD3/HIF Prolyl Hydroxylase 3 in 0.5 mg/mL of Hypoxic HeLa cell lysate. This experiment was performed under reducing conditions using the ...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 (NB100-303) at 1:3000 ...read more
Western Blot: EGLN3/PHD3 Antibody [NB100-303] - Tamoxifen administration-induced PHD2 deletion in skeletal muscle in Phd2f/f/Rosa26CreERT2 mice. Anti-PHD2 and PHD3 Western blotting of gastrocnemius and soleus muscles at ...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 X-100. The cells were ...read more
Immunohistochemistry: EGLN3/PHD3 Antibody [NB100-303] - PHD3 was detected in immersion fixed paraffin-embedded sections of human heart using Rabbit Anti-Human EGLN3 polyclonal Antibody (NB100-303) at 5 ug/mL for 1 hour ...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.

Product Details

Summary
Reactivity Hu, Mu, Rt, Pm, RtSpecies Glossary
Applications WB, Simple Western, ChIP, IB, ICC/IF, IHC, 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
  • Immunohistochemistry-Paraffin
  • Immunoprecipitation 1:10-1:500
Application Notes
In Western blot, a band is seen at approx. 27 kDa. This antibody has been tested against HeLa and MEF cell lysates. Immunoprecipitation was reported in scientific literature (PMID: 15721254). Chromatin Immunoprecipitation was reported in scientific literature (PMID: 21620138). Use in Immunohistochemistry-Paraffin reported in scientific literature (PMID: 25161887). Use in Immunocytochemistry/immunofluorescence reported in scientific literature (PMID: 20801873). In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
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.
Control
HeLa Whole Cell Lysate
EGLN3/PHD3 Overexpression Lysate
EGLN3/PHD3 Knockout HeLa Cell Lysate
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
    2 publications
  • WB
    25 publications

Reactivity Notes

Primate reactivity reported in scientific literature (PMID: 23732909). Rat reactivity reported in scientific literature (PMID: 23545307).

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, Rt, Pm, Rt
Applications: WB, Simple Western, ChIP, IB, ICC/IF, IHC, IHC-P, IP

Publications for EGLN3/PHD3 Antibody (NB100-303)(40)

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.


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ChIP
(1)
IB
(1)
ICC/IF
(1)
IHC-P
(1)
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(2)
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(25)
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Human
(15)
Mouse
(15)
Rat
(1)
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Showing Publications 1 - 10 of 40. Show All 40 Publications.
Publications using NB100-303 Applications Species
Xu YM, Gao Q, Zhang JZ et al. Prolyl hydroxylase 3 controls the intestine goblet cell generation through stabilizing ATOH1 Cell Death Differ. Jan 20 2020 [PMID: 31959916] (WB, IP, Human, Mouse) WB, IP Human, Mouse
Xu Y, Gao Q, Xue Y et al. Prolyl hydroxylase 3 stabilizes the p53 tumor suppressor by inhibiting the p53-MDM2 interaction in a hydroxylase-independent manner J. Biol. Chem. May 15 2019 [PMID: 31092600] (WB, Human) WB Human
Egners A, Rezaei M, Kuzmanov A et al. PHD3 Acts as Tumor Suppressor in Mouse Osteosarcoma and Influences Tumor Vascularization via PDGF-C Signaling Cancers (Basel) Dec 6 2018 [PMID: 30563292] (WB, Mouse) WB Mouse
Chowdhury A, Aich A, Jain G et al. Defective Mitochondrial Cardiolipin Remodeling Dampens HIF-1a Expression in Hypoxia. Cell Rep. Oct 16 2018 [PMID: 30332638] (Mouse) Mouse
Yano H, Sakai M, Matsukawa T et al. PHD3 regulates glucose metabolism by suppressing stress-induced signalling and optimising gluconeogenesis and insulin signalling in hepatocytes. Sci Rep Sep 24 2018 [PMID: 30250231] (WB, Mouse, Human) WB Mouse, Human
Sim J, Cowburn AS, Palazon A et al. The Factor Inhibiting HIF Asparaginyl Hydroxylase Regulates Oxidative Metabolism and Accelerates Metabolic Adaptation to Hypoxia. Cell Metab. Apr 3 2018 [PMID: 29617647] (WB, Mouse) WB Mouse
Bialesova L, Xu L, Gustafsson JA et al. Estrogen receptor beta 2 induces proliferation and invasiveness of triple negative breast cancer cells: association with regulation of PHD3 and HIF-1 alpha Oncotarget 2017 Sep 29 [PMID: 29100336] (WB, Human) WB Human
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
Show All 40 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.

Reviews using NB100-303:
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IF
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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. 

Control Lysate(s)

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Alexa Fluor 350 NB100-303AF350
Alexa Fluor 405 NB100-303AF405
Alexa Fluor 488 NB100-303AF488
Alexa Fluor 594 NB100-303AF594
Alexa Fluor 647 NB100-303AF647
Alexa Fluor 700 NB100-303AF700
Alexa Fluor 750 NB100-303AF750
Biotin NB100-303B
DyLight 350 NB100-303UV
DyLight 405 NB100-303V
DyLight 488 NB100-303G
DyLight 550 NB100-303R
DyLight 594 NB100-303DL594
DyLight 650 NB100-303C
DyLight 680 NB100-303FR
DyLight 755 NB100-303IR
FITC NB100-303F
HRP NB100-303H
Janelia Fluor 549 NB100-303JF549
Janelia Fluor 646 NB100-303JF646

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Recent Reviews

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2
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1
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Helder Andre
05/14/2015
Application: IF
Species: Human

Bioinformatics

Gene Symbol EGLN3
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