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.

Western Blot: EGLN1/PHD2 Antibody [NB100-138] - Detection of Human PHD2 by Western Blot. Samples: Recombinant epitope-tagged PHD1, PHD2 or PHD3 (10 ng/lane) or whole cell lysate from HeLa cells, stained with rabbit anti-PHD2 used at 1 ug/ml.

• Reactivity: Hu, Rt, Mu(-)
• Applications: WB, EM, Flow, ICC/IF, IHC, IHC-P, IP, KD
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1 mg/ml

EGLN1/PHD2 Antibody Summary

• Immunogen: Synthetic peptide made 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:
  • Western Blot 1:500-1:2500
  • Electron Microscopy
  • Flow Cytometry 1:10-1:1000
  • Immunocytochemistry/Immunofluorescence
  • Immunohistochemistry
  • Immunohistochemistry-Paraffin
  • Immunoprecipitation
  • Knockdown Validated
• 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 7.0 - 8.0)
• Preservative: 0.09% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for EGLN1/PHD2 Antibody

• C1orf12
• C1orf12HPH2
• EC 1.14.11
• EC 1.14.11.-
• ECYT3
• egl nine homolog 1 (C. elegans)
• egl nine homolog 1
• egl nine-like protein 1
• EGLN1
• HIF prolyl hydroxylase 2
• HIFPH2 EGL nine (C.elegans) homolog 1
• HIFPH2
• HIF-PH2
• HIF-prolyl hydroxylase 2
• HPH2
• HPH-2
• Hypoxia-inducible factor prolyl hydroxylase 2
• PHD2
• PHD2SM20
• Prolyl hydroxylase domain-containing protein 2
• SM20
• SM-20
• zinc finger MYND domain-containing protein 6
• ZMYND6
• ZMYND6DKFZp761F179

Background

HIF prolyl 4-hydroxylases (PHDs) are proyl hydroxylase domain-containing enzymes (PHD1/ Egln2, PHD2/ Egln1, PHD3/ Egln3, and P4H-TM) that mediate important physiological responses to hypoxia by modulating HIF1alpha levels. The HIF-alpha 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). Under normoxic conditions, HIF Prolyl Hydroxylase 2 (HIF1-PH; also called PHD1 or Egln2) catalyzes the post-translational formation of 4-hydroxyproline through hydroxylation of a specific proline found in each of NODD/CODD domains of HIF1A and also hydroxylates HIF2A with preference for CODD site of HIF1A/HIF1B. After hydroxylation, HIFs undergo proteasomal degradation via VHL (von Hippel-Lindau) ubiquitination complex. However, under hypoxic conditions, the hydroxylation reaction is tempered which allows HIFs to escape degradation process followed by their nuclear translocation, heterodimerization with HIF1B, and increased expression of hypoxia-inducible genes. HIF1-PH, through regulating HIF1 stability, is involved in various hypoxia-influenced processes such as angiogenesis in retinal/cardiac functionality as well as tumor angiogenesis, and defecticve EGLN1 have been linked to ECYT3 (familial erythrocytosis type 3).

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)(14)

Publications using NB100-138

• Tan J, Prosser H, Dunn L et al. High-Density Lipoproteins Rescue Diabetes-Impaired Angiogenesis via Scavenger Receptor Class B Type I. Diabetes Jul 8 1905 [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 May. [PMID: 21877141] (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 [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 Jun 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 [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 [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 Oct 2 [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 [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 [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 Dec 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 Oct [PMID: 17003483] (WB, IHC-P, ICC/IF, EM, Rat)
• Ietta, F, Wu, Y, Winter, J, Xu, J, Wang, J, Post, M, Caniggia, I. (2006) Dynamic HIF1A Regulation During Human Placental Development. Biol Reprod75(1):112-21. 2006 Jul. [PMID: 16611863]
• Cappuzzo F, Toschi L, Tallini G, Ceresoli GL et al. 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 Oncol;17(7):1120-7. 2006 Jul. [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 Feb 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: Anonymous WB Human 10/04/2015

Summary

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

Blocking

• Blocking Details: Blocking with 5% non-fat milk for 30 min

Primary Anitbody

• Dilution Ratio: 1:1000, overnight

Secondary Antibody

• Secondary Description: Donkey anti-rabbit IgG, HRP
• Secondary Manufacturer Cat#: GE Healthcare, NA934V
• Secondary Concentration: 1:2000

Details

• Detection Notes: Regular Western blot Protocol

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

1. What is the meaning of mouse(negative)? Can it be used in mouse?
   • The listing "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.

Bioinformatics

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