EGLN1/PHD2 Antibody

Immunocytochemistry/Immunofluorescence: EGLN1/PHD2 Antibody [NB100-2219] - Analysis of EGLN1/PHD2 in ARPE-19 cells using anti-PHD2 antibody. Image from veriifed customer review.

Immunohistochemistry-Paraffin: EGLN1/PHD2 Antibody [NB100-2219] - Analysis of a FFPE mouse lung section using 1:200 dilution of EGLN1/PHD2 antibody. The staining was developed using HRP conjugated anti-rabbit secondary antibody and DAB reagent. The antibody generated a specific staining in the cytoplasm and nuclei of alveolar as well as bronchiolar epithelial cells. Cytoplasmic staining was observed in almost all cells while the nuclear positivity was seen in a subset of cells only.

Immunocytochemistry/Immunofluorescence: EGLN1/PHD2 Antibody [NB100-2219] - EGLN1/PHD2 antibody at 1:500 in HeLa cells with DyLight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and DyLight 550 (red).

Western Blot: EGLN1/PHD2 Antibody [NB100-2219] - Detection of EGLN1/PHD2 in mouse kidney lysate. ECL exposure, 20 seconds.

Immunohistochemistry: EGLN1/PHD2 Antibody [NB100-2219] - Staining of renal tubular epithelium in mouse using NB100-2219 at 2.5 ug/mL.

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

EGLN1/PHD2 Antibody Summary

• Immunogen: A synthetic peptide made to an internal portion of mouse PHD2/HIF Prolyl Hydroxylase 2 (between residues 300-400). [Uniprot: Q91YE3]
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: EGLN1
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Western Blot 2 ug/mL
  • Immunocytochemistry/Immunofluorescence 1:50 - 1:500
  • Immunohistochemistry 2.5 - 5.0 ug/mL
  • Immunohistochemistry-Paraffin 2.5 - 5.0 ug/ml
  • Immunoprecipitation 1:10 - 1:500
  • Knockout Validated
• Application Notes: In Western blot a band is seen ~43 kDa representing HIF Prolyl Hydroxylase 2. There is also a non-specific band of similar intensity at ~75 kDa.
  
  Knockout Validation was reported in scientific literature (PMID: 24695462).
• Theoretical MW: 43 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:

  Mouse Kidney Whole Tissue Lysate (Adult Whole Normal)

Packaging, Storage & Formulations

• Storage: Store at 4C. Do not freeze.
• Buffer: PBS
• Preservative: 0.02% 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 (NB100-2219)(31)

Publications using NB100-2219

• Kim S, Jo CH, Kim GH Effects of empagliflozin on nondiabetic salt-sensitive hypertension in uninephrectomized rats Hypertens. Res. Sep 19 2019 [PMID: 31537914] (WB, Rat)
• Chowdhury D, Alrefai H, Landero Figueroa JA et al. Metallothionein 3 Controls the Phenotype and Metabolic Programming of Alternatively Activated Macrophages Cell Rep Jun 25 2019 [PMID: 31242420] (WB, Mouse)
• Yamamoto A, Hester J, Macklin PS, et al. Systemic silencing of PHD2 causes reversible immune regulatory dysfunction J. Clin. Invest. 2019 Jun 04 [PMID: 31162141] (WB, Mouse)
• Sim J, Cowburn AS, Palazon A et al. The Factor Inhibiting HIF Asparaginyl Hydroxylase Regulates Oxidative Metabolism and Accelerates Metabolic Adaptation to Hypoxia. Cell Metab. Apr 3 2018 [PMID: 29617647] (WB, Mouse)
• 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)
• Flisinski M, Wisniewska-Chudy E, Brymora A et al. Chronic kidney disease leads to hypoxia inducible factor-1alpha to hypoxia inducible factor-2alpha switch in the gastrocnemius muscle J. Physiol. Pharmacol. 2017 Jun 01 [PMID: 28820398] (WB, Rat)
• Wang S, Zeng H, Chen ST et al. Ablation of endothelial prolyl hydroxylase domain protein-2 promotes renal vascular remodelling and fibrosis in mice. J. Cell. Mol. Med. Mar 7 2017 [PMID: 28266128] (WB, Mouse)
• Choi JE, Kim JH, Song NY et al. 15-Deoxy-delta12, 14-prostaglandin J2 stabilizes hypoxia inducible factor-1alpha through induction of heme oxygenase-1 and direct modification of prolyl-4-hydroxylase 2. Free Radic. Res. Sep 6 2016 [PMID: 27598034] (IP, Human)
• Zeng H, He X, Tuo QH et al. LPS causes pericyte loss and microvascular dysfunction via disruption of Sirt3/angiopoietins/Tie-2 and HIF-2alpha/Notch3 pathways. Sci Rep. 2016 Feb 12 [PMID: 26868537] (WB, Human, Mouse)
• Karsikas S, Myllymaki M, Heikkila M et al. HIF-P4H-2 deficiency protects against skeletal muscle ischemia-reperfusion injury. J Mol Med (Berl) 2016 Mar [PMID: 26452676]
• Xie L, Pi X, Townley-Tilson WH et al. PHD2/3-dependent hydroxylation tunes cardiac response to beta-adrenergic stress via phospholamban. J Clin Invest 2015 Jul 1 [PMID: 26075818] (WB)
• Wu S, Zhang J, Huang C et al. Prolyl hydroxylase domain-2 silencing induced by hydrodynamic limb vein injection enhances vascular regeneration in critical limb ischemia mice through activation of multiple genes. Curr Gene Ther 2015 [PMID: 25832622] (Mouse)
• Hesse AR, Levent E, Zieseniss A et al. Lights on for HIF-1a: Genetically Enhanced Mouse Cardiomyocytes for Heart Tissue Imaging. Cell. Physiol. Biochem. 2014 Jul 30 [PMID: 25095893] (WB, Mouse)
  
  Details:
  PHD2/HIF Prolyl Hydroxylase 2 antibody used for WB on protein extracts of hypoxia exposed ODD-Luc-CMs/Cardiomyocytes derived from an embryonic stem cell (ESC) lines (ODD-Luc ESCs) that was generated from a Tg ROSA26 ODD-Luc/+ mouse (Figure 2D).
• Zimmermann AS, Morrison SD, Hu MS et al. Epidermal or dermal specific knockout of PHD-2 enhances wound healing and minimizes ischemic injury. PLoS One 2014 Apr 2 [PMID: 24695462] (KO, Mouse)
• Swain Lija, Wottawa Marieke, Hillemann Annette et al. Prolyl-4-hydroxylase domain 3 (PHD3) is a critical terminator for cell survival of macrophages under stress conditions. J Leukoc Biol. 2014 Mar 13 [PMID: 24626957] (Mouse)
• An X, Xu G, Yang L et al. Expression of hypoxia-inducible factor-1alpha, vascular endothelial growth factor and prolyl hydroxylase domain protein 2 in cutaneous squamous cell carcinoma and precursor lesions and their relationship with histological stages and clinical features. J Dermatol 2013 Dec 20 [PMID: 24354513] (IHC-P, Human)
• Kim J, Kwak HJ, Cha JY et al. The role of prolyl hydroxylase domain protein (PHD) during rosiglitazone-induced adipocyte differentiation. J Biol Chem 2013 Dec 12 [PMID: 24338020] (WB, Mouse)
• Palomaki S, Pietila M, Laitinen S et al. HIF-1alpha is upregulated in human mesenchymal stem cells. Stem Cells 2013 Sep [PMID: 23744828] (WB, Human)
• Pisarcik S, Maylor J, Lu W et al. Activation of Hypoxia-Inducible Factor-1 in Pulmonary Arterial Smooth Muscle Cells by Endothelin-1. Am J Physiol Lung Cell Mol Physiol 2013 Feb 15 [PMID: 23418090] (WB, Rat)
• Kunze R, Zhou W, Veltkamp R et al. Neuron-specific prolyl-4-hydroxylase domain 2 knockout reduces brain injury after transient cerebral ischemia. Stroke 2012 Oct [PMID: 22933585] (Mouse)
• Kuzmanov A, Wielockx B, Rezaei M et al. Overexpression of factor inhibiting HIF-1 enhances vessel maturation and tumor growth via platelet-derived growth factor-C. International Journal of Cancer 2011 Nov 18 [PMID: 22095574]
• Tandon R, SenGupta D, Ndhlovu LC et al. Identification of human endogenous retrovirus-specific T cell responses in vertically HIV-1-infected subjects. J Virol 2011 Nov [PMID: 21880743] (WB)
• Rabie T, Kunze R, Marti HH. Impaired hypoxic response in senescent mouse brain. Int J Dev Neurosci. 2011 Jun 16. [PMID: 21704147]
• 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]
• Wu S, Nishiyama N, Kano MR et al. Enhancement of angiogenesis through stabilization of hypoxia-inducible factor-1 by silencing prolyl hydroxylase domain-2 gene. Mol Ther 2008 Jul [PMID: 18500250] (WB)
• Kariuki TM, Van Dam GJ, Deelder AM et al. Previous or ongoing schistosome infections do not compromise the efficacy of the attenuated cercaria vaccine. Infect Immun 2006 Jul [PMID: 16790771]
• Haag D. Rolle der Siah-Ubiquitinligasen in der Entstehung der pulmonalen Hypertonie und daraus resultierenden rechtsventrikularen Hypertrophie. Einleitung. 2014 (WB, Mouse)
• Hoelscher M, Silter M, Krull S et al. Cardiomyocyte-specific Prolyl-4-hydroxylase domain 2 knock out protects from acute myocardial ischaemic injury. J Biol Chem. 2011 [PMID: 21270129]
• Hyvarinen J, Hassinen IE, Sormunen R et al. Hearts of hypoxia-inducible factor prolyl 4-hydroxylase-2 hypomorphic mice show protection against acute ischemia-reperfusion injury. J Biol Chem. 2010 Apr 30 [PMID: 20185832] (WB, Mouse)
• Schodel J, Klanke B, Weidemann A et al. HIF-Prolyl Hydroxylases in the Rat Kidney: Physiologic Expression Patterns Regulation in Acute Kidney Injury. Am J Pathol;174(5):1663-1674. 2009 [PMID: 19349364] (WB, Mouse)
• Mikhaylova O, Ignacak ML, Barankiewicz TJ et al. The von Hippel-Lindau tumor suppressor protein and Egl-9-Type proline hydroxylases regulate the large subunit of RNA polymerase II in response to oxidative stress. Mol Cell Biol;28(8):2701-17. 2008 Apr. [PMID: 18285459] (WB, Human)

Reviews for EGLN1/PHD2 Antibody (NB100-2219) (3)

Average Rating: 4.7

(Based on 3 reviews)

We have 3 reviews tested in 2 species: Human, Mouse.

reviewed by: Anonymous WB Mouse 02/16/2017

Summary

• Application: Western Blot
• Sample Tested: murine bone marrow derived macrophages
• Species: Mouse

reviewed by: Helder Andre IF Human 05/14/2015

Summary

• Application: Immunofluorescence
• Sample Tested: ARPE-19 cells
• Species: Human

Blocking

• Blocking Details: 10% goat serum in PBS-TritonX (0.1%) (30min @ RT)

Primary Anitbody

• Dilution Ratio: 1:500 in blocking solution (1h @ RT)

Secondary Antibody

• Secondary Description: 1:1000 Goat anti-rabbit IgG-A488 in blocking solution (1h @ RT)

Details

• Detection Notes: Epifluorescence microscope
• Fixation Details: Fixed in 4% buffered formalin (15min @ RT); Permeabilized in PBS-TritonX (0.5%) (10min @ RT)
• Wash Description: 3x5min PBS

reviewed by: Anonymous WB Mouse 01/18/2012

Summary

• Application: Western Blot
• Sample Tested: nuclear extract (brain), Sample Amount: 20ug
• Species: Mouse

Blocking

• Blocking Details: Blocking Buffer: 10% TPBS, Blocking Time: 1 hour, Blocking Temp: room temperature

Primary Anitbody

• Dilution Ratio: Dilution Ratio: 1:1000, Incubation Dilution Buffer: blocking buffer, Incubation Time: overnight, Incubation Temp: 4 C

Bioinformatics

• Gene Symbol: EGLN1