EGLN3/PHD3 Antibody (EG188e/d5)

Images

 
Western Blot: EGLN3/PHD3 Antibody (EG188e/d5) [NBP1-30440] - PHD3 expression in SK-N-BE(2) whole cell lysates. Cells were treated in normoxia (left) and in hypoxia (right). Image from verified customer review.
Immunohistochemistry: EGLN3/PHD3 Antibody (EG188e/d5) [NBP1-30440] - Analysis of HIF Prolyl Hydroxylase 3 in human renal cancer using DAB with hematoxylin counterstain.
Western Blot: EGLN3/PHD3 Antibody (EG188e/d5) [NBP1-30440] - Analysis of HIF Prolyl Hydroxylase 3 in Lane 1: Cobalt chloride treated COS-7 nuclear extracts and Lane 2: Untreated COS-7 nuclear extracts.
Simple Western: EGLN3/PHD3 Antibody (EG188e/d5) [NBP1-30440] - 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 ...read more

Product Details

Summary
Reactivity Hu, Rt, Pm, Mu, BvSpecies Glossary
Applications WB, Simple Western, IHC, IHC-P
Clone
EG188e/d5
Clonality
Monoclonal
Host
Mouse
Conjugate
Unconjugated
Concentration
1 mg/ml
   

Biological Strategies

     

EGLN3/PHD3 Antibody (EG188e/d5) Summary

Immunogen
Partial recombinant human PHD3/HIF Prolyl Hydroxylase 3 (UniProt# Q9H6Z9)
Localization
Cytoplasm. Nucleus.
Predicted Species
Mouse (98%), Bovine (98%). Backed by our 100% Guarantee.
Isotype
IgG1
Clonality
Monoclonal
Host
Mouse
Gene
EGLN3
Purity
Protein G 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 10 ug/ml
  • Simple Western 1:1000
  • Immunohistochemistry 1:400
  • Immunohistochemistry-Paraffin 1:400
Application Notes
This HIF Prolyl Hydroxylase 3 antibody is useful for Immunohistochemistry paraffin embedded sections and Western blot, where a band is seen approx. 27 kDa. Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (pH 6.0) is recommended.

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.
Reviewed Applications
Read 1 Review rated 4
using
NBP1-30440 in the following applications:

Publications
Read Publications using
NBP1-30440 in the following applications:

  • 1 publication
  • WB
    3 publications

Reactivity Notes

Human, monkey and rat. Immunogen sequence has 98% identity to bovine and mouse.

Packaging, Storage & Formulations

Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Buffer
Tris-Glycine and 0.15M NaCl
Preservative
0.05% Sodium Azide
Concentration
1 mg/ml
Purity
Protein G purified

Alternate Names for EGLN3/PHD3 Antibody (EG188e/d5)

  • 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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Publications for EGLN3/PHD3 Antibody (NBP1-30440)(5)

We have publications tested in 2 confirmed species: Human, Rat.

We have publications tested in 2 applications: IHC-P, WB.


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IHC-P
(1)
WB
(3)
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Human
(3)
Rat
(1)
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Review for EGLN3/PHD3 Antibody (NBP1-30440) (1) 41

Average Rating: 4
(Based on 1 review)
We have 1 review tested in 1 species: Human.
We have 1 review tested in 1 application: WB.

Reviews using NBP1-30440:
Filter by Applications
WB
(1)
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Human
(1)
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Images Ratings Applications Species Date Details
Western Blot EGLN3/PHD3 NBP1-30440
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reviewed by:
Anonymous
WB Human 11/20/2015
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Summary

ApplicationWestern Blot
Sample TestedMCF-7 whole cell lysate
SpeciesHuman
LotB-1

Blocking

Blocking DetailsPBST with 5% non-fat dry milk, 1 h, Room temperature

Primary Anitbody

Dilution Ratio10 ug/ml

Secondary Antibody

Secondary DescriptionGoat anti-mouse IgG HRP
Secondary Manufacturer Cat#sc-2005
Secondary Concentration1:2000

Details

Detection NotesChemiluminescence, G Box Chemi XL1.4 from Syngene, 5 mins, no poitive or negative controls

Comments

CommentsWash blots few extra times before developing.

Product General Protocols

View specific protocols for EGLN3/PHD3 Antibody (NBP1-30440): Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol

FAQs for EGLN3/PHD3 Antibody (NBP1-30440). (Showing 1 - 1 of 1 FAQs).

  1. I received an inquiry about Anti-PHD3/HIF Prolyl Hydroxylase 3 Antibody (EG188e/d5), #NBP1-30440. Could you let me know if this antibody cross-reacts with PHD1 or PHD2?
    • We have not tested our PHD3 antibody for cross reactivity with PHD1 or PHD2, therefore we are unaware of any possible cross reactivity.

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Alexa Fluor 405 Labeled NBP1-30440AF405
Alexa Fluor 488 Labeled NBP1-30440AF488
Alexa Fluor 647 Labeled NBP1-30440AF647
Alexa Fluor 700 Labeled NBP1-30440AF700
Biotin Labeled NBP1-30440B
DyLight 405 Labeled NBP1-30440V
DyLight 488 Labeled NBP1-30440G
DyLight 550 Labeled NBP1-30440R
DyLight 650 Labeled NBP1-30440C
FITC Labeled NBP1-30440F
HRP Labeled NBP1-30440H

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Recent Reviews

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1
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Anonymous
11/20/2015
Application: WB
Species: Human

Bioinformatics

Gene Symbol EGLN3
Entrez
Uniprot