HIF Prolyl Hydroxylases Antibody Pack

Simple Western: HIF Prolyl Hydroxylases Antibody Pack [NB100-903PHD] - Simple Western lane view shows a specific band for PHD2/HIF Prolyl Hydroxylase 2 in 0.5 mg/mL of Hypoxic HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system. EGLN1/PHD2 Antibody [NB100-137]

Knockdown Validated: HIF Prolyl Hydroxylases Antibody Pack [NB100-903PHD] - PHD3 is required for SCC cell survival in prolonged hypoxia. PHD3 is induced in hypoxia. Knock-down of PHD3 expression with siRNA (siPHD3) is specific and does not affect PHD2 or HIF-1 alpha expression. siScr = scrambled control siRNA. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0014617), licensed under a CC-BY licence. EGLN3/PHD3 Antibody [NB100-139]

Immunohistochemistry: HIF Prolyl Hydroxylases Antibody Pack [NB100-903PHD] - Immunofluorescence of wounded tissue and normal unwounded skin. Representative images for immunofluorescent staining of PHD-2 on healed wounded skin and adjacent normal unwounded skin of wild type, heterozygous K14-Cre/homozygous floxed PHD-2, and heterozygous Col1 alpha2-Cre-ER/homozygous floxed PHD-2 mice at 400x magnification. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0093373), licensed under a CC-BY licence. EGLN1/PHD2 Antibody [NB100-2219]

Western Blot: HIF Prolyl Hydroxylases Antibody Pack [NB100-903PHD] - Whole cell protein from MEF cells was separated on a 12% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2 ug/mL anti-EGLN3/PHD3 in 1% milk, and detected with an anti-rabbit HRP secondary antibody using chemiluminescence. EGLN3/PHD3 Antibody [NB100-303]

Immunocytochemistry: HIF Prolyl Hydroxylases Antibody Pack [NB100-903PHD] - EGLN1/PHD2 Antibody [NB100-137] - Staining of endogenous PHD2 in U2OS cells. ICC image submitted by a verified customer review. EGLN1/PHD2 Antibody [NB100-137]

Immunohistochemistry: HIF Prolyl Hydroxylases Antibody Pack [NB100-903PHD] - PHD3 was detected in immersion fixed paraffin-embedded sections of human heart using Rabbit Anti-Human EGLN3 polyclonal Antibody (NB100-303) at 5 ug/mL for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte(TM) HRP Polymer Antibody (VC003). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to nuclei. EGLN3/PHD3 Antibody [NB100-303]

Flow Cytometry: HIF Prolyl Hydroxylases Antibody Pack [NB100-903PHD] - Detection of PHD2. 1 million Jurkat cells were fixed, permeabilized, and stained with 3.0 ug/mL anti-PHD2 NB100-137 in a 150 uL reaction. Isotype control (black), anti-MLL1 (red).

Western Blot: HIF Prolyl Hydroxylases Antibody Pack [NB100-903PHD] - 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. EGLN1/PHD2 Antibody [NB100-138]

Western Blot: HIF Prolyl Hydroxylases Antibody Pack [NB100-903PHD] - Whole cell lysate (50 ug) from HeLa and 293T cells prepared using NETN lysis buffer. Antibody: Affinity purified rabbit anti-PHD4 antibody used for WB at 0.4 ug/mL. Detection: Chemiluminescence with an exposure time of 30 seconds. PHD4/HIF Prolyl Hydroxylase 4 Antibody [NB100-295]

Western Blot: HIF Prolyl Hydroxylases Antibody Pack [NB100-903PHD] - Analysis of PHD2 expression in human prostate cancer cell lines. WB image submitted by a verified customer review. EGLN1/PHD2 Antibody [NB100-137]

Immunohistochemistry: HIF Prolyl Hydroxylases Antibody Pack [NB100-903PHD] - 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. EGLN1/PHD2 Antibody [NB100-2219]

Immunohistochemistry: HIF Prolyl Hydroxylases Antibody Pack [NB100-903PHD] - Analysis of a formalin fixed tissue section of human renal cell carcinoma (clear cell type) using rabbit polyclonal EGLN3/PHD3 antibody with HRP-DAB detection and hematoxylin counterstaining. The antibody generated a strong nuclear staining of PHD3 primarily in the cancer cells while the stromal cells were largely negative for this protein. EGLN3/PHD3 Antibody [NB100-139]

Knockdown Validated: HIF Prolyl Hydroxylases Antibody Pack [NB100-903PHD] - In vitro analysis of PHD-2 knockout and protein quantification. Western blot data for PHD-2 knockout in fibroblasts of heterozygous Col1 alpha2-Cre-ER/homozygous floxed PHD-2 mice compared to wild type mice (*p<0.05). D') Western blot data for PHD-2 knockout in keratinocytes of heterozygous K14-Cre/homozygous floxed PHD-2 mice compared to wild type mice (*p<0.05). Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0093373), licensed under a CC-BY licence. EGLN1/PHD2 Antibody [NB100-2219]

• Reactivity: Hu, Mu, Rt
• Applications: WB, Simple Western, Flow, ICC/IF, IHC, IHC-P, ICC/IF, KD
• Host: Rabbit
• Concentration: Concentration of individual antibodies may be found on the vial label. If unlisted please contact technical services.

HIF Prolyl Hydroxylases Antibody Pack Summary

• Immunogen: The exact amino acid range is proprietary. General Immunogen area:
  NB100-137 (LocusLink - 54583 Region - N-Terminus),
  NB100-138 (LocusLink - 54583 Region - C-Terminus),
  NB100-139 (LocusLink - 112399 Region - Internal),
  NB100-295 (LocusLink - 54681 Region - C-terminus),
  NB100-303 (LocusLink - 112399 Region - C-terminus),
  NB100-2219 (mouse amino acid range 300-400).
• Specificity: See individual datasheets.
• Host: Rabbit
• Gene: EGLN1
• Purity: Immunogen affinity purified

Applications/Dilutions

• Application Notes: These antibodies can be used for Western blot at a dilution range of 1:500 - 1:1,000. Other applications are not confirmed.
  See individual datasheets for validated applications.
• Control:

  EGLN1/PHD2 Overexpression Lysate

Reactivity Notes

All: Homo sapiens.
NB100-137 does not react with mouse.
NB100-138 does not react with mouse and mixed results in rat.
NB100-139 does not react with mouse but reacts with rat.
NB100-295 does not react with mouse and is unknown in rat.
NB100-303 reacts with mouse.
Other species have not been tested.

Packaging, Storage & Formulations

• Storage: Store at 4C. Do not freeze.
• Preservative: 0.01% Sodium Azide
• Concentration: Concentration of individual antibodies may be found on the vial label. If unlisted please contact technical services.
• Purity: Immunogen affinity purified

Notes

This pack contains 1 vial each of: NB100-2219SS (0.025 mL), NB100-303SS (0.025 mL), NB100-139SS (0.025 mL), NB100-138SS (0.025 mL), NB100-295SS (0.025 mL), NB100-137SS (0.025 mL).

Alternate Names for HIF Prolyl Hydroxylases Antibody Pack

• 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
• HIF prolyl hydroxylase 2
• HIF-PH2
• HIFPH2EGL nine (C.elegans) homolog 1
• HIF-prolyl hydroxylase 2
• HPH-2
• Hypoxia-inducible factor prolyl hydroxylase 2
• PHD2SM20
• Prolyl hydroxylase domain-containing protein 2
• SM-20
• zinc finger MYND domain-containing protein 6
• ZMYND6DKFZp761F179

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Antibody Packs are guaranteed for 1 year from date of receipt.

Publications for HIF Prolyl Hydroxylases Antibody Pack (NB100-903PHD)(15)

Publications using NB100-903PHD

• Siddiq A, Aminova LR, Troy CM et al. Selective Inhibition of Hypoxia-Inducible Factor (HIF) Prolyl-Hydroxylase 1 Mediates Neuroprotection against Normoxic Oxidative Death via HIF- CREB-Independent Pathways. J Neurosci;29(27):8828-8838. 2009 [PMID: 19587290]
• Loinard C, Ginouves A, Vilar J et al. Inhibition of Prolyl Hydroxylase Domain Proteins Promotes Therapeutic Revascularization. Circulation;120(1):50-59. 2009 [PMID: 19546390]
• Schultz K, Murthy V, Tatro JB et al. Prolyl hydroxylase 2 deficiency limits proliferation of vascular smooth muscle cells by hypoxia-inducible factor-1{alpha}-dependent mechanisms. Am J Physiol Lung Cell Mol Physiol;296(6):L921-927. 2009 [PMID: 19304911]
• Mikhaylova, O, Ignacak, M 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]
• Chang, E et al. Age decreases endothelial progenitor cell recruitment through decreases in hypoxia-inducible factor 1alpha stabilization during ischemia. Circulation;116(24):2818-29. 2007 Dec 11. [PMID: 18040029]
• Bex, C et al. A yeast two-hybrid system reconstituting substrate recognition of the von Hippel-Lindau tumor suppressor protein. Nucleic Acids Res;35(21):e142. 2007 [PMID: 17986458]
• Baek, JH et al. Spermidine/spermine N(1)-acetyltransferase-1 binds to hypoxia-inducible factor-1alpha (HIF-1alpha) and RACK1 and promotes ubiquitination and degradation of HIF-1alpha. J Biol Chem;282(46):33358-66. 2007 Nov 16. [PMID: 17875644]
• Colla S, Tagliaferri S, Morandi F 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]
• Baek, JH et al. Spermidine/spermine-N1-acetyltransferase 2 is an essential component of the ubiquitin ligase complex that regulates hypoxia-inducible factor 1alpha. J Biol Chem;282(32):23572-80. 2007 Aug 10. [PMID: 17558023]
• Choi, Y et al. AKAP12 regulates human blood-retinal barrier formation by downregulation of hypoxia-inducible factor-1alpha. J Neurosci;27(16):4472-81. 2007 Apr 18. [PMID: 17442832]
• Grover, TR et al. Hypoxia-inducible factors HIF-1alpha and HIF-2alpha are decreased in an experimental model of severe respiratory distress syndrome in preterm lambs. Am J Physiol Lung Cell Mol Physiol;292(6):L1345-51. 2007 Jun. [PMID: 17307811]
• Kondo, S et al. EBV latent membrane protein 1 up-regulates hypoxia-inducible factor 1alpha through Siah1-mediated down-regulation of prolyl hydroxylases 1 and 3 in nasopharyngeal epithelial cells. Cancer Res;66(20):9870-7. 2006 Oct 15. [PMID: 17047048]
• McMahon, S et al. Transforming growth factor beta1 induces hypoxia-inducible factor-1 stabilization through selective inhibition of PHD2 expression. J Biol Chem;281(34):24171-81. 2006 Aug 25. [PMID: 16815840]
• Stiehl, DP et al. Increased prolyl 4-hydroxylase domain proteins compensate for decreased oxygen levels. Evidence for an autoregulatory oxygen-sensing system. J Biol Chem;281(33):23482-91. 2006 Aug 18. [PMID: 16790428]
• Lu H et al. Reversible inactivation of HIF-1 prolyl hydroxylases allows cell metabolism to control basal HIF-1. J Biol Chem;280(51):41928-39. 2005 Dec 23. [PMID: 16223732]

Bioinformatics

• Gene Symbol: EGLN1