EGLN1/PHD2 Antibody

Western Blot: EGLN1/PHD2 Antibody [NB100-138] - Analysis of human PHD2, using NB100-138. Samples: Recombinant FLAG-His-PHD1, PHD2 and PHD3 (10 ng/lane), HeLa whole cell lysate and MEFs.

Flow Cytometry: EGLN1/PHD2 Antibody [NB100-138] - Flow cytometric detection of PHD2, 10^6 Jurkat cells were fixed, permeabilized, and stained with 3.0 ug/mL anti-PHD2 in a 150 uL reaction.

Product Discontinued

EGLN1/PHD2 Antibody Summary

• Immunogen: This EGLN1/PHD2 antibody was developed against a synthetic peptide corresponding to a C-terminal portion of human PHD2/HIF Prolyl Hydroxylase 2 (between amino acids 350-426) (GeneID 54583).
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: EGLN1
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Electron Microscopy
  • Flow Cytometry 1:10 - 1:1000
  • Immunocytochemistry/ Immunofluorescence
  • Immunohistochemistry
  • Immunohistochemistry-Paraffin
  • Immunoprecipitation
  • Knockdown Validated
  • Western Blot 1:500 - 1:2500
• Application Notes: This PHD2/HIF Prolyl Hydroxylase 2 antibody is useful for Flow Cytometry and Western blot, where a band can be seen at 46-50 kDa. Immunoprecipitation and Immunohistochemistry were reported in scientific literature. Use in Immunocytochemistry/immunofluorescence, Immunohistochemistry-Paraffin and Electron Microscopy reported in scientific literature (PMID: 17003483). 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

Reactivity Notes

Does not appear to work in mouse. Rat reactivity reported in scientific literature (PMID: 17003483).

Packaging, Storage & Formulations

• Storage: Store at 4C. Do not freeze.
• Buffer: Tris-Citrate/Phosphate (pH 7 - 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.

Publications for EGLN1/PHD2 Antibody (NB100-138)(15)

Publications using NB100-138

• Bur H, Haapasaari K M et al. Strong Prolyl Hydroxylase Domain 1 Expression Predicts Poor Outcome in Radiotherapy-treated Patients with Classical Hodgkin's Lymphoma. Anticancer Res 2018-01-01 [PMID: 29277791] (IF/IHC, Human)
• Tan J, Prosser H, Dunn L et al. High-Density Lipoproteins Rescue Diabetes-Impaired Angiogenesis via Scavenger Receptor Class B Type I. Diabetes 1905-07-08 [PMID: 27284113]
• Peurala E, Koivunen P, Bloigu R, Haapasaari KM, Jukkola-Vuorinen A. Expressions of individual PHDs associate with good prognostic factors and increased proliferation in breast cancer patients. Breast Cancer Res Treat;133(1):179-88. 2012-05-01 [PMID: 21877141] (IF/IHC, Human)
• Mikheyev AS, Vo T, Wee B et al. Rapid microsatellite isolation from a butterfly by de novo transcriptome sequencing: performance and a comparison with AFLP-derived distances. PLoS One 2010-01-01 [PMID: 20585453]
• Serra-Perez A, Planas AM, Nunez-O'Mara A et al. Extended ischemia prevents HIF1alpha degradation at reoxygenation by impairing prolyl-hydroxylation: role of Krebs cycle metabolites. J Biol Chem 2010-06-11 [PMID: 20368331] (WB, Human)
• Sakamoto T, Seiki M. Mint3 Enhances the Activity of Hypoxia-inducible Factor-1 (HIF-1) in Macrophages by Suppressing the Activity of Factor Inhibiting HIF-1. J Biol Chem;284(44):30350-30359. 2009-01-01 [PMID: 19726677]
• Lehmann S, Stiehl DP, Honer M et al. Longitudinal multimodal in vivo imaging of tumor hypoxia its downstream molecular events. PNAS;106(33):14004-14009. 2009-01-01 [PMID: 19666490]
• Steinhoff A, Pientka FK, Mockel S et al. Cellular oxygen sensing: Importins and exportins are mediators of intracellular localisation of prolyl-4-hydroxylases PHD1 and PHD2. Biochem Biophys Res Commun 2009-10-02 [PMID: 19631610] (IP, Human)
• Xie L, Xiao K, Whalen EJ et al. Oxygen-Regulated {beta}2-Adrenergic Receptor Hydroxylation by EGLN3 Ubiquitylation by pVHL. Sci Signal;2(78):ra33-. 2009-01-01 [PMID: 19584355]
• Wong W, Goehring AS, Kapiloff MS et al. mAKAP Compartmentalizes Oxygen-Dependent Control of HIF-1{alpha}. Sci Signal;1(51):ra18-. 2008-01-01 [PMID: 19109240]
• Colla S, Tagliaferri S, Morandi F, Lunghi P, Donofrio G et al. The new tumor-suppressor gene inhibitor of growth family member 4 (ING4) regulates the production of proangiogenic molecules by myeloma cells and suppresses hypoxia-inducible factor-1 alpha (HIF-1alpha) activity: involvement in myeloma-induced angiogenesis. Blood;110(13):4464-75. 2007-12-15 [PMID: 17848618]
• Khan Z, Michalopoulos GK, Stolz DB. Peroxisomal localization of hypoxia-inducible factors and hypoxia-inducible factor regulatory hydroxylases in primary rat hepatocytes exposed to hypoxia-reoxygenation. Am J Pathol 2006-10-01 [PMID: 17003483] (IHC-P, ICC/IF, EM, WB, Rat)
• Ietta, F et al. Dynamic HIF-1alpha regulation during human placental development. Biol Reprod. 2006-01-01 [PMID: 16611863]
• Cappuzzo, F, Toschi, L, Tallini, G, Ceresoli, G L, Domenichini, I, Bartolini, S, Finocchiaro, G, Magrini, E, Metro, G, Cancellieri, A, Trisolini, R, Crino, L, Bunn, P A, Jr, Santoro, A, Franklin, W A, Varella-Garcia, M, Hirsch, F R. Insulin-like growth factor receptor 1 (IGFR-1) is significantly associated with longer survival in non-small-cell lung cancer patients treated with gefitinib. Ann Oncol17(7):1120-7. 2006-07-01 [PMID: 16600976]
• Baek JH, Mahon PC, Oh J et al. OS-9 interacts with hypoxia-inducible factor 1alpha and prolyl hydroxylases to promote oxygen-dependent degradation of HIF-1alpha. Mol Cell;17(4):503-12. 2005-02-18 [PMID: 15721254] (IP, Human)

Review for EGLN1/PHD2 Antibody (NB100-138) (1)

Average Rating: 4

(Based on 1 review)

We have 1 review tested in 1 species: Human.

reviewed by: Verified Customer WB Human 10/04/2015

Summary

• Application: Western Blot
• Sample Tested: Human cancer cell whole cell lysate
• Species: Human

FAQs for EGLN1/PHD2 Antibody (NB100-138). (Showing 1 - 2 of 2 FAQs).

1. What is the meaning of "mouse (negative)"? Can the antibody be used in mouse?
   • "Mouse (negative)" indicates that this antibody has been tested with samples from mouse, and found not to work. NB100-138 is covered by our 100% quality guarantee for use with samples from human and rat.
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.

Bioinformatics

• Gene Symbol: EGLN1
• Entrez:
  • 54583(Human)
• Uniprot:
  • Q9GZT9(Human)