EGLN1/PHD2 Antibody (2445B)

Western Blot: EGLN1/PHD2 Antibody (2445B) [NBP2-76810] - Total protein from human HeLa, THP-1 and U87 cell lines 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 2.0 ug/ml anti-PHD2 in block buffer and detected with an anti-rabbit HRP secondary antibody using West Pico PLUS chemiluminescence detection reagent.

Immunohistochemistry-Paraffin: EGLN1/PHD2 Antibody (2445B) [NBP2-76810] - EGLN1/PHD2 was detected in immersion fixed paraffin-embedded sections of human kidney using Rabbit Anti-EGLN1/PHD2 Monoclonal Antibody (Catalog #NBP2-76810) at 3 ug/mL for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell cytoplasm and nuclei.

Immunocytochemistry/Immunofluorescence: EGLN1/PHD2 Antibody (2445B) [NBP2-76810] - EGLN1/PHD2 was detected in immersion fixed WM-115 human malignant melanoma cell line using Rabbit Anti-EGLN1/PHD2 Monoclonal Antibody (Catalog #NBP2-76810) at 3 ug/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to cell cytoplasm and nuclei.

Flow Cytometry: EGLN1/PHD2 Antibody (2445B) [NBP2-76810] - Human Jurkat T Cell Leukemia Cell Line was stained with Rabbit Anti- EGLN1/PHD2 Monoclonal Antibody (Catalog # NBP2-76810, filled histogram) or Rabbit IgG Isotype Control Antibody (Catalog # MAB1050, open histogram) followed by Phycoerythrin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0110). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012).

• Reactivity: Hu
• Applications: WB, Flow, Flow-IC, ICC/IF, IHC, IHC-P
• Clone: 2445B
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1.0 mg/ml

EGLN1/PHD2 Antibody (2445B) Summary

• Additional Information: Recombinant Monoclonal Antibody
• Immunogen: Partial synthetic peptide from the N terminal portion of the human EGLN1/PHD2 protein
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Gene: EGLN1
• Purity: Protein A or G purified

Applications/Dilutions

• Dilutions:
  • Western Blot 1 - 2 ug/ml
  • Flow Cytometry 0.25 ug
  • Immunocytochemistry/Immunofluorescence 5 - 20 ug/ml
  • Immunohistochemistry 5 - 20 ug/ml
  • Immunohistochemistry-Paraffin 5 - 20 ug/ml
  • Flow (Intracellular) 0.25 ug

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Protein A or G purified

Alternate Names for EGLN1/PHD2 Antibody (2445B)

• 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 hydroxylase 2 is a prolyl hydroxylase that modifies HIF-alpha. Classic prolyl hydroxylases are found in the endoplasmic reticulum and modify collagen, whereas HIF is an intracellular protein and the HPH sites do not resemble those modifying collagen. HIF is a transcriptional complex that plays a critical role in oxygen homeostasis. HPH is an essential component of the pathway through which cells sense oxygen. In the presence of oxygen, HPHs convert specific prolyl residues in HIF-alpha to hydroxyproline, leading to HIF-alpha destruction. Low oxygen levels, sensed at the cellular level, cause the HIF conversion to be reduced so that HIF is stable and there is increased angiogenesis. HPH-2, specifically, catalyzes the posttranslational formation of 4-hydroxyproline in HIF alpha proteins. It hydroxylates HIF-1 alpha at Pro(402) and Pro(564), and HIF-2 alpha. It targets HIF through the hydroxylation for proteasomal degradation via the von Hippel-Lindau ubiquitylation complex.

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.

Bioinformatics

• Gene Symbol: EGLN1