EGLN3/PHD3 Antibody

Images

 
Biological Strategies: Knockdown Validated: EGLN3/PHD3 Antibody [NB100-139] - PHD3 is required for SCC cell survival in prolonged hypoxia. PHD3 is induced in hypoxia. Knock-down of PHD3 expression with siRNA ...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
Genetic Strategies: Knockdown Validated: EGLN3/PHD3 Antibody [NB100-139] - EGLN3/PHD3 depletion increases S10 phosphorylation of p27 in hypoxia. S10 phosphorylated form of p27 is strongly induced in EGLN3/PHD3 ...read more
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
Genetic Strategies: Knockdown Validated: EGLN3/PHD3 Antibody [NB100-139] - BxPC3 cells stably transduced with retrovirus containing PHD3Wt (BxPC3-Wt), Vector (BxPC3-Vec) or anti-PHD3 shRNA (BxPC3-KD) were ...read more
Western Blot: EGLN3/PHD3 Antibody [NB100-139] - The HIF-1 response is not impaired in prostate and breast carcinoma cell lines that lack PHD3. Melanoma, breast and prostate cancer cell lines with methylated PHD3 ...read more
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)

Product Details

Summary
Reactivity Hu, Mu, RtSpecies Glossary
Applications WB, EM, ICC/IF, IHC, IHC-P, IP, KD
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
  • Knockdown Validated
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.
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.
Control
EGLN3/PHD3 Overexpression Lysate
EGLN3/PHD3 Knockout HeLa Cell Lysate
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
    2 publications
  • WB
    22 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 prolyl hydroxylase domain-containing enzymes (PHD1/ Egln2, PHD2/ Egln1, PHD3/ Egln3, and P4H-TM) that are known for their role in mediating physiological responses to hypoxic stress via modulation of HIF-1 alpha expression levels. The HIF-1 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 HIF-1 alpha's NODD/CODD domains and also hydroxylates HIF-2 alpha 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 allows HIFs to escape the 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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NB100-139
Species: Hu, Mu, Rt
Applications: WB, EM, ICC/IF, IHC, IHC-P, IP, KD

Publications for EGLN3/PHD3 Antibody (NB100-139)(44)

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
(2)
WB
(22)
All Applications
Filter By Species
Human
(17)
Mouse
(5)
Rat
(6)
All Species
Showing Publications 1 - 10 of 44. Show All 44 Publications.
Publications using NB100-139 Applications Species
Shi M, Dai WQ, Jia RR et al. APCCDC20-mediated degradation of PHD3 stabilizes HIF-1a and promotes tumorigenesis in hepatocellular carcinoma Cancer Lett Oct 9 2020 [PMID: 33039559] (IP, WB, Mouse) IP, WB Mouse
Kozlova N, Mennerich D et al. The pro-oncogenic adaptor CIN85 acts as an inhibitory binding partner of hypoxia-inducible factor prolyl hydroxylase 2. Cancer Res May 29 2019 [PMID: 31142511] (WB, Human) WB Human
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
Kozlova N, Mennerich D, Samoylenko A et al. The pro-oncogenic adaptor CIN85 inhibits hypoxia-inducible factor prolyl hydroxylase-2. Diabetes. Nov 19 2018 (WB, Human) WB Human
Miikkulainen P, Hogel H, Seyednasrollah F et al. Hypoxia-inducible factor (HIF)-prolyl hydroxylase 3 (PHD3) maintains high HIF2A mRNA levels in clear cell renal cell carcinoma. J. Biol. Chem. Jan 7 2019 [PMID: 30617181] (WB, Human) WB Human
Pan H, Strickland A, Madhu V et al. RNA binding protein HuR regulates extracellular matrix gene expression and pH homeostasis independent of controlling HIF-1a signaling in nucleus pulposus cells Matrix Biol. Aug 6 2018 [PMID: 30092282] (WB) WB
Akat F, Ficicilar H, Durak A et al. Intermittent hypoxia induces beneficial cardiovascular remodeling in left ventricular function of type 1 diabetic rat. Anatol J Cardiol 2018 Apr [PMID: 29615543] (Rat) Rat
Bryant JD, Brown MC, Dobrikov MI et al. Regulation of HIF-1A during Hypoxia by DAP5-Induced Translation of PHD2. Mol. Cell. Biol. Mar 12 2018 [PMID: 29530922] (WB) WB
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
Show All 44 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.

Reviews using NB100-139:
Filter by Applications
WB
(2)
All Applications
Filter by Species
Human
(1)
Mouse
(1)
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Images Ratings Applications Species Date Details
Western Blot EGLN3/PHD3 NB100-139
Enlarge
5
reviewed by:
Anonymous
WB Mouse 05/08/2016
View

Summary

ApplicationWestern Blot
SpeciesMouse
FileView PDF
  5
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.

Control Lysate(s)

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Alexa Fluor 350 NB100-139AF350
Alexa Fluor 405 NB100-139AF405
Alexa Fluor 488 NB100-139AF488
Alexa Fluor 532 NB100-139AF532
Alexa Fluor 594 NB100-139AF594
Alexa Fluor 647 NB100-139AF647
Alexa Fluor 700 NB100-139AF700
Alexa Fluor 750 NB100-139AF750
Biotin NB100-139B
DyLight 350 NB100-139UV
DyLight 405 NB100-139V
DyLight 488 NB100-139G
DyLight 550 NB100-139R
DyLight 594 NB100-139DL594
DyLight 650 NB100-139C
DyLight 680 NB100-139FR
DyLight 755 NB100-139IR
FITC NB100-139F
HRP NB100-139H
Janelia Fluor 549 NB100-139JF549
Janelia Fluor 646 NB100-139JF646

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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