EGLN3/PHD3 Antibody

Images

 
Western Blot: EGLN3/PHD3 Antibody [NB100-139] - Analysis of HeLa lysates using NB100-139. Image courtesy of Dr Gregg Semenza (The Johns Hopkins University School of Medicine, Baltimore, MD USA)
Immunocytochemistry/ Immunofluorescence: EGLN3/PHD3 Antibody [NB100-139] - EGLN3/PHD3 antibody was tested in HeLa cells with Dylight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and Dylight ...read more
Immunohistochemistry-Paraffin: EGLN3/PHD3 Antibody [NB100-139] - Analysis of a formalin fixed tissue section of human renal cell carcinoma (clear cell type) using rabbit polyclonal EGLN3/PHD3 antibody with HRP-DAB ...read more
Western Blot: EGLN3/PHD3 Antibody [NB100-139] - Detection of PHD3 by Western Blot application in lysate from Rat cells before or 9 hours after inducing hypoxia. Primary antibody PHD3 NB100-139 was used at a dilution of ...read more
Immunohistochemistry-Paraffin: EGLN3/PHD3 Antibody [NB100-139] - Analysis of a formalin fixed tissue section of human renal cell carcinoma (clear cell type) using rabbit polyclonal EGLN3/PHD3 antibody with HRP-DAB ...read more

Product Details

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

EGLN3/PHD3 Antibody Summary

Immunogen
Synthetic peptide corresponding to residues between 50-100 of human PHD3/HIF Prolyl Hydroxylase 3 using the numbering given in entry NP_071356.1 (GeneID 112399).
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 1:1000-1:2000
  • Electron Microscopy
  • Immunocytochemistry/Immunofluorescence 1:500
  • Immunohistochemistry 1:250-1:1000
  • Immunohistochemistry-Paraffin 1:250-1:1000
  • Immunoprecipitation
Application Notes
In WB, PHD3 band can be seen at 27-30 kDa molecular weight range. Use of this antibody in Electron Microscopy and ChIP is reported in scientific literature - PMIDs 17003483and 24367580 respectively. 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
28 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
EGLN3/PHD3 Lysate (NBL1-10156)
Reviewed Applications
Read 2 Reviews rated 5
using
NB100-139 in the following applications:

Publications
Read Publications using
NB100-139 in the following applications:

  • EM
    1 publication
  • 3 publications
  • 2 publications
  • IP
    1 publication
  • WB
    14 publications

Reactivity Notes

Mouse reactivity reported in scientific literature (PMID: 24037093)

Packaging, Storage & Formulations

Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Buffer
PBS
Preservative
0.05% 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.

Customers Who Viewed This Item Also Viewed...

NB100-137
Species: Hu, Mu, Rt, Pm
Applications: WB, Simple Western, Flow, ICC/IF, IHC, IHC-P, IP
NB100-105
Species: Hu, Mu, Rt, Po, Bv, Ft, Mk, Pm, Rb, Sh, Xp, Ye
Applications: WB, ChIP, ELISA, Flow, GS, ICC/IF, IHC, IHC-Fr, IHC-P, IP, PLA, TCS, KO, LA
AF6394
Species: Hu
Applications: WB, IP
NB100-485
Species: Hu
Applications: WB, IP
NBP1-89843
Species: Hu
Applications: ICC/IF, IHC, IHC-P
NB100-122
Species: Hu, Mu, Rt, Fi, Ha, Pm, Sh
Applications: WB, Simple Western, ChIP, ELISA, Flow, GS, ICC/IF, IHC, IHC-Fr, IHC-P, IP, KO
NB100-124
Species: Hu, Mu, Rt, Bv, Ft, Pm, Sh
Applications: WB, ChIP, GS, ICC/IF, IHC, IHC-P, IP
NB100-428
Species: Hu, Mu, Rt
Applications: WB, EM, ICC/IF, IHC, IHC-Fr, IHC-P, IP
NB120-19347
Species: Hu, Mu, Rt, Po, Bv
Applications: WB, ELISA, Flow, ICC/IF, IHC, IHC-Fr, IHC-P, IF
NBP1-84949
Species: Hu
Applications: ICC/IF, IHC, IHC-P
NB300-500
Species: Hu, Mu, Rt, Po, Am, Bv, Ca, Dr
Applications: WB, IHC, IHC-Fr, IHC-P
NBP1-89150
Species: Hu, Mu, Rt
Applications: WB, IHC, IHC-P
H00011026-M01
Species: Hu
Applications: WB, ELISA
NBP1-31302
Species: Hu, Mu, Rt
Applications: WB, ICC/IF, IHC, IHC-P
NBP1-92132
Species: Hu
Applications: WB, ICC/IF, IHC, IHC-P
NB100-139
Species: Hu, Mu, Rt
Applications: WB, EM, ICC/IF, IHC, IHC-P, IP

Publications for EGLN3/PHD3 Antibody (NB100-139)(35)

We have publications tested in 3 confirmed species: Human, Mouse, Rat.

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


Filter By Application
EM
(1)
ICC/IF
(3)
IHC-P
(2)
IP
(1)
WB
(14)
All Applications
Filter By Species
Human
(12)
Mouse
(3)
Rat
(5)
All Species
Showing Publications 1 - 10 of 35. Show All 35 Publications.
Publications using NB100-139 Applications Species
Miikkulainen P, Hogel H, Rantanen K et al. HIF prolyl hydroxylase PHD3 regulates translational machinery and glucose metabolism in clear cell renal cell carcinoma Cancer Metab 2017 Jul 06 [PMID: 28680592] (WB, Human) WB Human
Bruno T, Valerio M, Casadei L et al. Che-1 sustains hypoxic response of colorectal cancer cells by affecting Hif-1alpha stabilization. J. Exp. Clin. Cancer Res. 2017 Feb 18 [PMID: 28214471] (WB, Human) WB Human
Oh ET, Kim JW, Kim JM et al. NQO1 inhibits proteasome-mediated degradation of HIF-1a. Nat Commun. Dec 14 2016 [PMID: 27966538] (Mouse) Mouse
Adam MG, Matt S, Christian S et al. SIAH ubiquitin ligases regulate breast cancer cell migration and invasion independent of the oxygen status. Cell Cycle. 2015 Dec 02 [PMID: 26654769] (WB, Human) WB Human
Iriondo O, Rabano M, Domenici G et al. Distinct breast cancer stem/progenitor cell populations require either HIF1alpha or loss of PHD3 to expand under hypoxic conditions. Oncotarget 2015 Oct 13 [PMID: 26372732] (WB) WB
Hogel H, Miikkulainen P, Bino L, Jaakkola PM. Hypoxia inducible prolyl hydroxylase PHD3 maintains carcinoma cell growth by decreasing the stability of p27. Mol. Cancer. 2015 Jul 30 [PMID: 26223520] (WB, ICC/IF, Human)

Details:
PHD3 antibody used for WB and ICC/IF on human samples. ICC/IF performed on siPHD3-depleted HeLa cells (Figure 2A). WB carried out on HeLa and 786-O cells that were transfected with PHD3 siRNA followed by synchronization at G0 and 24-h hypoxic exposure (Figure 1).
WB, ICC/IF Human
Li J, Yuan W, Jiang S et al. Prolyl-4-hydroxylase Domain Protein 2 Controls NF-kB/p65 Transactivation and Enhances the Catabolic Effects of Inflammatory Cytokines on cells of the Nucleus Pulposus J. Biol. Chem. 2015 Jan 29 [PMID: 25635047] (WB, Rat, Human)

Details:
EGLN3/PHD3 antibody used for WB on lysates from nucleus pulposus/NP cells (rat) that were transduced with Sh-control or Sh-PHD2 with or without TNF-alpha treatment (FIGURE 1-I). Antibody also used for WB analysis of immunoprecipitates obtained from HEK293T cells that were transfected with plasmids of FLAG-p65, YFP-PHD2 or PHD3 (FIGURE 6B)
WB Rat, Human
Vogler M, Zieseniss A, Hesse AR et al. Pre- and post-conditional inhibition of prolyl-4-hydroxylase domain enzymes protects the heart from an ischemic insult Pflugers Arch. 2015 Jan 13 [PMID: 25578858] (IHC-P, Mouse)

Details:
PHD3/HIF Prolyl Hydroxylase 3 antibody used at 1:10,000 dilution for IHC-P application on heart sections from C57BL/6 mice after 1, 6, and 24 h of treatment (40-fold magnification) with ICA, a specific PHD inhibitor 2-(1-chloro-4-hydroxyisoquinoline-3-carboxamido) acetate - two doses of 40 mg/kg BW ICA or vehicle i.p., either 6 and 1 h before myocardial infarction or 1 and 5 h after myocardial infarction (Fig 3).
IHC-P Mouse
Place TL, Nauseef JT, Peterson MK et al. Prolyl-4-Hydroxylase 3 (PHD3) Expression Is Downregulated during Epithelial-to-Mesenchymal Transition. PLoS One 2013 Dec 18 [PMID: 24367580] (WB, IP, Human) WB, IP Human
Rawluszko AA, Bujnicka KE, Horbacka K et al. Expression and DNA methylation levels of prolyl hydroxylases PHD1, PHD2, PHD3 and asparaginyl hydroxylase FIH in colorectal cancer. BMC Cancer. 2013 Nov 6 [PMID: 24195777] (WB, Human) WB Human
Show All 35 Publications.

Reviews for EGLN3/PHD3 Antibody (NB100-139) (2) 52

Average Rating: 5
(Based on 2 reviews)
We have 2 reviews tested in 2 species: Human, Mouse.
We have 2 reviews tested in 1 application: WB.

Reviews using NB100-139:
Filter by Applications
WB
(2)
All Applications
Filter by Species
Human
(1)
Mouse
(1)
All Species
Images Ratings Applications Species Date Details
Western Blot EGLN3/PHD3 NB100-139
Enlarge
reviewed by:
Anonymous
WB Mouse 05/08/2016
View

Summary

ApplicationWestern Blot
SpeciesMouse
FileView PDF
 
reviewed by:
Anonymous
WB Human 06/24/2010
View

Summary

ApplicationWestern Blot
Sample TestedHeLa cell lysates, Sample Amount: 20 ug
SpeciesHuman

Blocking

Blocking DetailsBlocking Buffer: 5% milk in TBST, Blocking Time: 1 hour, Blocking Temp: 25 degrees Celsius

Primary Anitbody

Dilution RatioPrimary Ab Dilution Ratio: 1:1000, Primary Ab Incubation Time: overnight, Primary Ab Incubation Temp: 4 degrees Celsius

Secondary Antibody

Secondary DescriptionSecondary Ab: donkey anti-rabbit IgG-HRP

Details

Detection NotesDetection Method: ECL Plus, Specific Bands: 27 kDa

Product General Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for EGLN3/PHD3 Antibody (NB100-139) (0)

There are no specific FAQs related to this product. Read our general customer & technical service FAQs.

Positive Control Lysate(s)

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Alexa Fluor 405 Labeled NB100-139AF405
Alexa Fluor 488 Labeled NB100-139AF488
Alexa Fluor 647 Labeled NB100-139AF647
Alexa Fluor 700 Labeled NB100-139AF700
Biotin Labeled NB100-139B
DyLight 405 Labeled NB100-139V
DyLight 488 Labeled NB100-139G
DyLight 550 Labeled NB100-139R
DyLight 650 Labeled NB100-139C
FITC Labeled NB100-139F
HRP Labeled NB100-139H

Additional EGLN3/PHD3 Products

Bioinformatics Tool for EGLN3/PHD3 Antibody (NB100-139)

Discover related pathways, diseases and genes to EGLN3/PHD3 Antibody (NB100-139). Need help? Read the Bioinformatics Tool Guide for instructions on using this tool.
Visit Tool

Diseases for EGLN3/PHD3 Antibody (NB100-139)

Discover more about diseases related to EGLN3/PHD3 Antibody (NB100-139).
 

Pathways for EGLN3/PHD3 Antibody (NB100-139)

View related products by pathway.

PTMs for EGLN3/PHD3 Antibody (NB100-139)

Learn more about PTMs related to EGLN3/PHD3 Antibody (NB100-139).
 

Research Areas for EGLN3/PHD3 Antibody (NB100-139)

Find related products by research area.

Blogs on EGLN3/PHD3

There are no specific blogs for EGLN3/PHD3, but you can read our latest blog posts.

Customers Who Bought This Also Bought

Contact Information

Product PDFs

Calculators

Concentration Calculator

The concentration calculator allows you to quickly calculate the volume, mass or concentration of your vial. Simply enter your mass, volume, or concentration values for your reagent and the calculator will determine the rest.

=
÷

Recent Reviews

5
5
2
4
0
3
0
2
0
1
0

Anonymous
05/08/2016
Application: WB
Species: Mouse

 
Anonymous
06/24/2010
Application: WB
Species: Human

Bioinformatics

Gene Symbol EGLN3
Entrez
Uniprot