EGLN1/PHD2 Antibody

Images

 
Western Blot: EGLN1/PHD2 Antibody [NB100-137] - Analysis of PHD2 expression in human prostate cancer cell lines. Western blot image submitted by a verified customer review.
Simple Western: EGLN1/PHD2 Antibody [NB100-137] - Simple Western lane view shows a specific band for PHD2/HIF Prolyl Hydroxylase 2 in 0.5 mg/mL of hypoxic HeLa cell lysate. This experiment was performed under reducing ...read more
Immunocytochemistry/ Immunofluorescence: EGLN1/PHD2 Antibody [NB100-137] - Staining of endogenous PHD2 in U2OS cells. ICC/IF image submitted by a verified customer review.
Genetic Strategies: Knockdown Validated: EGLN1/PHD2 Antibody [NB100-137] - Detection of PHD2 in human glioblastoma cell lysates. Image submitted by a verified customer review.
Western Blot: EGLN1/PHD2 Antibody [NB100-137] - Detection of human PHD2 by Western blot. Recombinant epitope-tagged PHD1, PHD2 or PHD3 (10 ng/lane) or whole cell lysate from HeLa cells. EGLN1/PHD2 antibody used at 1 ...read more
Western Blot: EGLN1/PHD2 Antibody [NB100-137] - Exogenous PHD2 has no impact on the gene expression of HIF-1alpha and HIF-2alpha. U87MG cells were transfected with empty vector (pcDNA) or PHD2 plasmid. Twenty-four hours ...read more
Immunohistochemistry: EGLN1/PHD2 Antibody [NB100-137] - EGLN1/PHD2 and EGFR expression levels positively correlate in breast cancer. Processed tissue microarrays of breast cancer biopsies from 313 patients were stained ...read more
Flow Cytometry: EGLN1/PHD2 Antibody [NB100-137] - An intracellular stain was performed on Jurkat cells with NB100-137AF647 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then ...read more
Western Blot: EGLN1/PHD2 Antibody [NB100-137] - Analysis of PHD2 in normoxia (lane 1) or hypoxia (lane 2) treated SK-N-BE(2) cells using anti-EGLN1/PHD2 antibody. Western blot image submitted by a verified customer ...read more
Immunocytochemistry/ Immunofluorescence: EGLN1/PHD2 Antibody [NB100-137] - Staining of PHD2 in HeLa cells with DyLight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and DyLight 550 (red).
Immunohistochemistry: EGLN1/PHD2 Antibody [NB100-137] - Staining of lung vascular endothelium with EGLN1/PHD2 antibody. Image at 40X.
Flow Cytometry: EGLN1/PHD2 Antibody [NB100-137] - Detection of PHD2 in Jurkat cells. One million Jurkat cells were fixed, permeabilized, and stained with 3.0 ug/mL anti-EGLN1/PHD2 antibody in a 150 uL reaction. Isotype ...read more

Product Details

Summary
Reactivity Hu, Mu, Rt, Pm, Pm-Or, Rb, RMSpecies Glossary
Applications WB, Simple Western, Flow, ICC/IF, IHC, IHC-P, IP, KD
Clonality
Polyclonal
Host
Rabbit
Conjugate
Unconjugated
Concentration
1 mg/ml
Validated by:
     

Genetic Strategies

   

EGLN1/PHD2 Antibody Summary

Immunogen
The epitope recognized by this EGLN1/PHD2 antibody maps to a region between residues 1 and 50 of human PHD2/HIF Prolyl Hydroxylase 2 using the numbering given in entry NP_071334.1 (GeneID 54583).
Localization
Cytoplasm, nucleus
Predicted Species
Rhesus Macaque (100%), Orangutan (100%), Rabbit (100%). Backed by our 100% Guarantee.
Isotype
IgG
Clonality
Polyclonal
Host
Rabbit
Gene
EGLN1
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 - 1:2500
  • Simple Western 1:500
  • Flow Cytometry 3.0 mcg/mL
  • Immunocytochemistry/Immunofluorescence 1:50
  • Immunohistochemistry 1:10 - 1:500
  • Immunohistochemistry-Paraffin 1:10 - 1:500
  • Immunoprecipitation
  • Knockdown Validated
Application Notes
This EGLN1/PHD2 antibody is useful for Flow Cytometry, Immunocytochemistry/Immunofluorescence, Western Blot, and Immunohistochemistry-paraffin embedded sections. In ICC/IF, cytoplamic and nuclear staining was observed in HeLa cells. Immunoprecipitation was reported in scientific literature.

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
46 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
EGLN1/PHD2 Overexpression Lysate
Reviewed Applications
Read 12 Reviews rated 4.8
using
NB100-137 in the following applications:

Publications
Read Publications using
NB100-137 in the following applications:

  • 5 publications
  • IHC
    6 publications
  • 4 publications
  • IP
    4 publications
  • WB
    49 publications

Reactivity Notes

Results for use of this EGLN1/PHD2 antibody have been mixed in Rat with success in Western blot analysis and immunofluorescence on Rat endothelial cells and negative results with PC12 cells. Mouse reactivity reported in scientific literature (PMID: 25578858). Rat reactivity reported in scientific literature (PMID: 25635047). Primate reactivity reported in scientific literature (PMID: 25974097)

Packaging, Storage & Formulations

Storage
Store at 4C. Do not freeze.
Buffer
Tris-Citrate/Phosphate (pH 7 to 8)
Preservative
0.09% Sodium Azide
Concentration
1 mg/ml
Purity
Immunogen affinity purified

Alternate Names for EGLN1/PHD2 Antibody

  • C1orf12
  • EC 1.14.11.29
  • EC:1.14.11.291
  • ECYT3
  • egl nine homolog 1
  • egl nine-like protein 1
  • EGLN1
  • HALAH
  • HIF prolyl hydroxylase 2
  • HIFPH2
  • HIF-PH2
  • HIF-prolyl hydroxylase 2
  • HPH2
  • HPH-2
  • Hypoxia-inducible factor prolyl hydroxylase 2
  • PHD2
  • PNAS-118
  • PNAS-137
  • Prolyl hydroxylase domain-containing protein 2
  • SM20
  • SM-20
  • zinc finger MYND domain-containing protein 6
  • ZMYND6

Background

PHD2 (Prolyl Hydroxylase Domain-containing protein 2) belongs to the Prolyl-4-hydroxylase domain (PHD) family of proteins and is encoded by the Egl-9 Family Hypoxia Inducible Factor 1 (EGLN1) gene (1). Human EGLN1/PHD2 is a ubiquitously expressed enzyme that is 426 amino acids (aa) long with a theoretical molecular weight of ~46 kDa. Structurally PHD2 contains a nuclear export signal (NES, aa 6-20), an N-terminal MYND zinc finger domain (aa 21-58), and a C-terminal catalytic domain (aa 291-392) (2, 3). Functionally, PHD2 serves as an oxygen sensor and is responsible for post-translational modification of Hypoxia-inducible factor alpha (HIF-1alpha), a component of a transcriptional complex involved in oxygen homeostasis (1-3). During normoxia, PHD2 is responsible for oxygen-dependent hydroxylation of HIF-1alpha proline residue 402, 564, or both (3). The hydroxylation event promotes the binding of von Hippel-Lindau protein (VHL) and targets HIF1-alpha for ubiquitination and degradation (4, 5).

EGLN1/PHD2 has been implicated in several critical processes including erythropoiesis, angiogenesis, and metabolism as well as various pathologies such as cancer (2, 5, 6). Studies in mice have found that somatic deletion of PHD2 resulted in higher vascular endothelial growth factor A (VEGF-A) levels, increased blood vessel formation, and more erythropoietin (EPO), leading to severe polycythemia or erythrocytosis (high red blood cell (RBC) volume) (6). Another study revealed that specific point mutations in EGLN1/PHD2 led to elevated EPO and RBC mass associated with hemorrhages and strokes (6). Accordingly, given the known role of PHD2 in inhibition of EPO production, PHD2 inhibitors are being studied as a potential therapeutic for anemia (6). Additionally, dysregulation in EGLN1, and specifically the PHD2-VHL-HIF-1alpha pathway, has been associated with the development of pheochromocytomas (PCC) and sympathetic paragangliomas (PGL), which are rare neuroendocrine tumors (2). Besides pathological features, EGLN1/PHD2 may also be important for high altitude adaptation as two coding sequence variants in PHD2 are prevalent in the Tibetan population but is very rare in people at lower altitudes (2).

Alternate names for EGLN1/PHD2 include HIF Prolyl Hydroxylase 2, PH2, Prolyl hydroxylase domain containing protein 2, HIF2PH2, HIF-Prolyl hydroxylase 2, egl nine homolog 1, and C1orf12.

References

1. Amorim-Pires, D., Peixoto, J., & Lima, J. (2016). Hypoxia Pathway Mutations in Pheochromocytomas and Paragangliomas. Cytogenetic and genome research. https://doi.org/10.1159/000457479

2. Gardie, B., Percy, M. J., Hoogewijs, D., Chowdhury, R., Bento, C., Arsenault, P. R., Richard, S., Almeida, H., Ewing, J., Lambert, F., McMullin, M. F., Schofield, C. J., & Lee, F. S. (2014). The role of PHD2 mutations in the pathogenesis of erythrocytosis. Hypoxia (Auckland, N.Z.). https://doi.org/10.2147/HP.S54455

3. Minervini, G., Quaglia, F., & Tosatto, S. C. (2015). Insights into the proline hydroxylase (PHD) family, molecular evolution and its impact on human health. Biochimie. https://doi.org/10.1016/j.biochi.2015.07.009

4. Semenza G. L. (2007). Hypoxia-inducible factor 1 (HIF-1) pathway. Science's STKE : signal transduction knowledge environment. https://doi.org/10.1126/stke.4072007cm8

5. Chan, D. A., & Giaccia, A. J. (2010). PHD2 in tumour angiogenesis. British journal of cancer. https://doi.org/10.1038/sj.bjc.6605682

6. Meneses, A. M., & Wielockx, B. (2016). PHD2: from hypoxia regulation to disease progression. Hypoxia (Auckland, N.Z.). https://doi.org/10.2147/HP.S53576

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 EGLN1/PHD2 Antibody (NB100-137)(78)

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

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


Filter By Application
ICC/IF
(5)
IHC
(6)
IHC-P
(4)
IP
(4)
WB
(49)
All Applications
Filter By Species
Human
(52)
Mouse
(5)
Rat
(7)
Primate
(1)
All Species
Showing Publications 1 - 10 of 78. Show All 78 Publications.
Publications using NB100-137 Applications Species
Bo H Role of oxygen on Nuclear factor erythroid-2-like 1 (NFE2L1 or Nrf1) function and stability Thesis (WB, Human) WB Human
Mei H, Wu N, Huang X et al. Possible mechanisms by which silkworm faeces extract ameliorates adenine-induced renal anaemia in rats J Ethnopharmacol Oct 3 2020 [PMID: 33022342] (WB, Rat) WB Rat
Settelmeier S, Schreiber T, MAki J et al. Prolyl hydroxylase domain 2 reduction enhances skeletal muscle tissue regeneration after soft tissue trauma in mice PLoS ONE May 15 2020 [PMID: 32413092] (IHC, Mouse) IHC Mouse
Dabral S, Muecke C, Valasarajan C et al. A RASSF1A-HIF1 alpha loop drives Warburg effect in cancer and pulmonary hypertension Nat Commun May 13 2019 [PMID: 31086178] (IP, Human) IP Human
Kurelac I, Iommarini L, Vatrinet R et al. Inducing cancer indolence by targeting mitochondrial Complex I is potentiated by blocking macrophage-mediated adaptive responses Nat Commun Feb 22 2019 [PMID: 30796225] (WB, Human) WB Human
Liu S, Zhang G, Guo J et al. Loss of Phd2 cooperates with BRAFV600E to drive melanomagenesis Nat Commun Dec 21 2018 [PMID: 30575721] (IHC, Human) IHC Human
Deygas M, Gadet R, Gillet G et al. Redox regulation of EGFR steers migration of hypoxic mammary cells towards oxygen. Nat Commun. Oct 31 2018 [PMID: 30382089] (WB, Human) WB Human
Kim G, Das R, Rao X et al. CITED2 restrains pro-inflammatory macrophage activation and response. Mol Cell Biol 2017 Dec 04 [PMID: 29203644] (WB, Mouse) WB Mouse
Hu J, Wang W, Zhang F et al. Hypoxia inducible factor-1 alpha mediates the profibrotic effect of albumin in renal tubular cells Sci Rep 2017 Nov 20 [PMID: 29158549] (WB, Rat) WB Rat
Bhute V, Harte J, Houghton J, Maxwell P Mannose binding lectin is hydroxylated by collagen prolyl-4-hydroxylase and inhibited by some PHD inhibitors Kidney360 Apr 3 2020 (WB, Human) WB Human
Show All 78 Publications.

Reviews for EGLN1/PHD2 Antibody (NB100-137) (12) 4.812

Average Rating: 4.8
(Based on 12 reviews)
We have 12 reviews tested in 3 species: Human, Human and mouse liver cells, Other.

Reviews using NB100-137:
Filter by Applications
WB
(10)
IF
(1)
All Applications
Filter by Species
Human
(9)
Human and mouse liver cells
(1)
Other
(2)
All Species
Images Ratings Applications Species Date Details
 EGLN1/PHD2 NB100-137
Enlarge
5
reviewed by:
Anonymous
Human and mouse liver cells 01/19/2020
View

Summary

Sample TestedProtein and paraffin sections
SpeciesHuman and mouse liver cells
LotA1
Western Blot EGLN1/PHD2 NB100-137
Enlarge
5
reviewed by:
John Collins
WB Human 10/23/2017
View

Summary

ApplicationWestern Blot
Sample TestedHuman epithelial cell lines
SpeciesHuman
LotA3
Western Blot EGLN1/PHD2 NB100-137
Enlarge
5
reviewed by:
Anonymous
WB Human 10/08/2015
View

Summary

ApplicationWestern Blot
Sample TestedOPM2 cells whole cell lysate
SpeciesHuman
LotA3

Blocking

Blocking DetailsPBST 0.2%Tween 20, 5% non fat milk, 30 minutes at Room Temperature

Primary Anitbody

Dilution Ratio1:1000, Overnight, 5 C, PBST with 5%milk

Secondary Antibody

Secondary DescriptionGoat anti-rabbit igG HRP
Secondary Manufacturer Cat#sc-2004
Secondary Concentration1:2000

Details

Detection NotesChemiluminescence, G:Box Chemi XLT4, 20 sec, no positive and negative controls

Comments

CommentsSemi-dry transfer method, used 4-12% Nupage gels from Life technologies
Western Blot EGLN1/PHD2 NB100-137
Enlarge
5
reviewed by:
Anonymous
WB Human 06/30/2015
View

Summary

ApplicationWestern Blot
Sample TestedCell lysates from prostate cancer cell lines
SpeciesHuman
LotLot A3

Blocking

Blocking Details4% BSA in TBST for 1h at room temperature

Primary Anitbody

Dilution Ratio1:500 in TBST + 1%BSA, overnight, 4*C

Secondary Antibody

Secondary DescriptionGoat anti-rabbit IgG-HRP
Secondary Manufacturer Cat#sc-2031
Secondary Concentration1:10,000

Details

Detection NotesECL method using Femto reagent
  4
reviewed by:
Anonymous
WB Human 01/05/2015
View

Summary

ApplicationWestern Blot
Sample TestedSee PMID 23225569
SpeciesHuman

Blocking

Blocking DetailsSee PMID 23225569

Details

Detection NotesSee PMID 23225569

Comments

CommentsPublished PMID 23225569
  4
reviewed by:
Anonymous
WB Human 12/12/2014
View

Summary

ApplicationWestern Blot
Sample TestedSee PMID 23328582
SpeciesHuman

Blocking

Blocking DetailsSee PMID 23328582

Details

Detection NotesSee PMID 23328582

Comments

CommentsPublished in PMID: 23328582Independent review from antibodyresource.com
  5
reviewed by:
Anonymous
WB Human 11/21/2014
View

Summary

ApplicationWestern Blot
SpeciesHuman
FileView PDF
  5
reviewed by:
Anonymous
WB Other 09/22/2014
View

Summary

ApplicationWestern Blot
Sample TestedHuman SH-SY5Y or rat PC12 whole cell lysate.
SpeciesOther
LotA

Blocking

Blocking Details5% milk in PBST for 1 hour at room temperature

Primary Anitbody

Dilution Ratio1:1000 in 1% BSA at 4oC overnight

Secondary Antibody

Secondary DescriptionGoat anti rabbit IgG, HRP
Secondary Manufacturer Cat#Cell Signaling #7074
Secondary Concentration1:2000

Details

Detection NotesECL 3 seconds
  5
reviewed by:
Anonymous
WB Other 08/24/2014
View

Summary

ApplicationWestern Blot
Sample TestedSK-N-BE(2) whole cell lysate
SpeciesOther
LotA

Blocking

Blocking Details5% milk in PBST for 1 hour at room temperature

Primary Anitbody

Dilution Ratio1:1000 in 1% BSA at 4oC overnight

Secondary Antibody

Secondary DescriptionGoat anti rabbit IgG, HRP
Secondary Manufacturer Cat#Cell Signaling #7074
Secondary Concentration1:2000

Details

Detection NotesECL 3 seconds
Immunofluorescence EGLN1/PHD2 NB100-137
Enlarge
5
reviewed by:
Anonymous
IF Human 04/28/2014
View

Summary

ApplicationImmunofluorescence
Sample TestedU2OS (human osteosarcoma cell line)
SpeciesHuman

Blocking

Blocking Details4% FSG (fish skin gelatin)/PBS, 4 degrees celsius, over night

Primary Anitbody

Dilution Ratio1:100 in 2% FSG/PBS, 4 degrees celsius, over night

Secondary Antibody

Secondary Descriptionanti-rabbit IgG, Alexa594
Secondary Manufacturer Cat#Invitrogen
Secondary Concentration1:800

Details

Detection NotesFluorescence microscope (Zeiss, Axioplan)
Fixation Details15Min room temperature 3.7% Formaldehyde, 10Min room temperature 0.1% Triton X-100
Wash DescriptionPBS, 5Min, three times
Western Blot EGLN1/PHD2 NB100-137
Enlarge
5
reviewed by:
Anonymous
WB Human 04/28/2014
View

Summary

ApplicationWestern Blot
Sample TestedU87MG, U138MG, U343MG whole cell lysates
SpeciesHuman

Blocking

Blocking Details1h, 3% milk/PBS, room temperature

Primary Anitbody

Dilution Ratio2h, 1:500 in 3% milk/PBS, room temperature

Secondary Antibody

Secondary DescriptionGoat anti-rabbit IgG, HRP
Secondary Manufacturer Cat#Dako
Secondary Concentration1:2000

Details

Detection NotesECL, X-ray film, 2 Min exposure
  5
reviewed by:
Anonymous
WB Human 07/08/2010
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 RatioDilution Ratio: 1:1000, Incubation Dilution Buffer: 5% Milk in TBST, Incubation Time: overnight, Incubation Temp: 4 C

Secondary Antibody

Secondary DescriptionSecondary Ab: donkey anti-rabbit IgG-HRP, Secondary Ab Dilution Ratio: 1:5000

Details

Detection NotesDetection Method: ECL Plus, Specific Bands: 46 kDa, Non-Specific Bands: 70, 80, 120 kDa

Product General Protocols

View specific protocols for EGLN1/PHD2 Antibody (NB100-137): 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 EGLN1/PHD2 Antibody (NB100-137). (Showing 1 - 2 of 2 FAQs).

  1. What is the probability of this antibody cross-reacting with the bovine protein. Can you send me the alignment score of the immunogen with the bovine protein?
    • The immunogen used to generate this antibody shares 100% homology with bovine. This antibody will likely cross react with bovine, however we have not yet tested it in that species. If you would be interested in testing this species, please take a look at our Innovators Reward Program.
  2. I would like to use this antibody but it has not been validated in my species of interest. Is there any way I can find out if it will work?
    • We offer risk-free testing of all of our primary antibodies. Please check out our Innovator's Reward Program and test this EGLN1-PHD2 antibody in any unvalidated species or application, without the financial risk of failure.

Control Lysate(s)

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Alexa Fluor 405 NB100-137AF405
Alexa Fluor 488 NB100-137AF488
Alexa Fluor 532 NB100-137AF532
Alexa Fluor 647 NB100-137AF647
Alexa Fluor 700 NB100-137AF700
Biotin NB100-137B
DyLight 350 NB100-137UV
DyLight 405 NB100-137V
DyLight 488 NB100-137G
DyLight 550 NB100-137R
DyLight 594 NB100-137DL594
DyLight 650 NB100-137C
DyLight 680 NB100-137FR
DyLight 755 NB100-137IR
FITC NB100-137F
HRP NB100-137H
Janelia Fluor 549 NB100-137JF549
Janelia Fluor 646 NB100-137JF646
PE NB100-137PE

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Bioinformatics Tool for EGLN1/PHD2 Antibody (NB100-137)

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Diseases for EGLN1/PHD2 Antibody (NB100-137)

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PTMs for EGLN1/PHD2 Antibody (NB100-137)

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Recent Reviews

4.8
5
10
4
2
3
0
2
0
1
0

Anonymous
01/19/2020
Application:
Species: Human and mouse liver cells

John Collins
10/23/2017
Application: WB
Species: Human

Anonymous
10/08/2015
Application: WB
Species: Human

Bioinformatics

Gene Symbol EGLN1
Entrez
Uniprot