Human/Rat EGLN1/PHD2 Antibody

Recombinant Monoclonal Antibody
Catalog # Availability Size / Price Qty
MAB7680-100
MAB7680-SP
Detection  of Human and Rat EGLN1/PHD2 by Western Blot.
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Human/Rat EGLN1/PHD2 Antibody Summary

Species Reactivity
Human, Rat
Specificity
Detects human EGLN/PHD2 in direct ELISAs. Detects human and rat EGLN/PHD2 in Western blots. In direct ELISAs, no cross-reactivity with human PHD1 and PHD3 is observed.
Source
Recombinant Monoclonal Rabbit IgG Clone # 2445B
Purification
Protein A or G purified from cell culture supernatant
Immunogen
S. frugiperda insect ovarian cell line Sf 21-derived recombinant human EGLN1/PHD2
Ala2-Phe426
Accession # Q9GZT9
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
1 µg/mL
See below
Flow Cytometry
0.25 µg/106 cells
See below
Immunohistochemistry
3-25 µg/mL
See below
CyTOF-ready
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
 
Immunocytochemistry
3-25 µg/mL
See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Data Examples

Western Blot Detection  of Human and Rat EGLN1/PHD2 by Western Blot. View Larger

Detection of Human and Rat EGLN1/PHD2 by Western Blot. Western blot shows lysates of HEK293T human embryonic kidney cell line, PC‑3 human prostate cancer cell line, and L6 rat myoblast cell line. PVDF membrane was probed with 1 µg/mL of Rabbit Anti-Human/Rat EGLN1/PHD2 Monoclonal Antibody (Catalog # MAB7680) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for EGLN1/PHD2 at approximately 49 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Flow Cytometry Detection of EGLN1/PHD2 in Human Jurkat cell line by Flow Cytometry. View Larger

Detection of EGLN1/PHD2 in Human Jurkat cell line by Flow Cytometry. Human Jurkat T Cell Leukemia Cell Line was stained with Rabbit Anti-Human/Rat EGLN1/PHD2 Monoclonal Antibody (Catalog # MAB7680, 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). View our protocol for Staining Membrane-associated Proteins.

Immunocytochemistry EGLN1/PHD2  in WM-115 Human Cell Lines. View Larger

EGLN1/PHD2 in WM-115 Human Cell Lines. EGLN1/PHD2 was detected in immersion fixed WM‑115 human malignant melanoma cell line using Rabbit Anti-Human/Rat EGLN1/PHD2 Monoclonal Antibody (Catalog # MAB7680) at 3 µg/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. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Immunohistochemistry EGLN1/PHD2  in Human Kidney. View Larger

EGLN1/PHD2 in Human Kidney. EGLN1/PHD2 was detected in immersion fixed paraffin-embedded sections of human kidney using Rabbit Anti-Human/Rat EGLN1/PHD2 Monoclonal Antibody (Catalog # MAB7680) at 3 µg/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. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: EGLN1/PHD2

PHD2 (Prolyl Hydroxylase Domain-containing protein 2; also HPH2, EGLN1 and HIF-PH2) is a 45-47 kDa dioxygenase member of the PH family of enzymes. It is ubiquitously expressed, and serves to regulate the availability of the oxygen-sensitive HIF transcription factor. Active HIF1 alpha is a heterodimer of alpha - and beta -subunits and when intact, promotes VEGF and EPO production. The beta -subunit is constitutively expressed, while alpha -subunit levels are regulated by intracellular oxygen concentration. At normoxic levels, the alpha -subunit is hydroxylated on Pro by one of three PHDs, inducing its ubiquitination/degradation. The hydroxylation event requires oxygen, and thus PH activity (particularly PHD2) is a measure of a cell's oxygen concentration. Human PHD2 is 426 amino acids (aa) in length. It contains an NES (aa 6-20), a Zn-finger region (aa 21-58), and a catalytic domain (aa 291-392). There are five nitrosylated cysteines plus one acetylated alanine. Two isoform variants are known, one that shows a deletion of aa 338-359, and another that contains a 17 aa substitution for aa 58-175. Over aa 157-426, human PHD2 shares 93% aa sequence identity with mouse PHD2.

Long Name
Egl Nine Homolog 1/Prolyl Hydroxylase Domain-containing Protein 2
Entrez Gene IDs
54583 (Human); 112405 (Mouse); 308913 (Rat)
Alternate Names
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

Product Datasheets

FAQs

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Cell and Tissue Staining Kits

Immunohistochemistry Reagents

Isotype Controls

Normal Rabbit IgG Control

Ctrl,CyTOF-ready

Reconstitution Buffers

Secondary Antibodies

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