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Human EGLN2/PHD1 Antibody

Catalog #: AF6394
7 Citations Datasheet / COA / SDS
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AF6394
AF6394-SP
Detection of Human EGLN2/PHD1 by Western Blot.
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Citations (7)
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Human EGLN2/PHD1 Antibody Summary

Species Reactivity
Human
Specificity
Detects human EGLN2/PHD1 in direct ELISAs and Western blots.
Source
Polyclonal Sheep IgG
Purification
Antigen Affinity-purified
Immunogen
S. frugiperda insect ovarian cell line Sf 21-derived recombinant human EGLN2/PHD1
Asp2-Thr407
Accession # Q96KS0
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Applications

Recommended Concentration
Sample
Western Blot
1 µg/mL
See below
Immunoprecipitation
25 µg/mL
Conditioned cell culture medium spiked with recombinant human EGLN2/PHD1

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.

Scientific Data

Western Blot Detection of Human EGLN2/PHD1 antibody by Western Blot. View Larger

Detection of Human EGLN2/PHD1 by Western Blot. Western blot shows lysates of MDA-MB-231 human breast cancer cell line. PVDF Membrane was probed with 1 µg/mL of Human EGLN2/PHD1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6394) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for EGLN2/PHD1 at approximately 48 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.

Immunohistochemistry Detection of Mouse EGLN2/PHD1 by Immunohistochemistry View Larger

Detection of Mouse EGLN2/PHD1 by Immunohistochemistry NQO1 promotesin vivo tumour growth.(a) RKO/pNQO1, RKO/shCont and RKO/pshNQO1 cells were injected subcutaneously into the right flank of athymic, 7-week-old female BALB/C nude mice, and tumour growth was assessed. Tumour volume (TV) was calculated by using the following formula: TV=length × (width)2 × 0.5. Each group contained 12 animals. (**P<0.01 with unpaired t-test). (b) Immunohistochemical analyses of RKO/pNQO1, RKO/shCont, and RKO/pshNQO1 xenograft tumours. The sections were stained for NQO1, HIF-1 alpha, proliferation (Ki67) and apoptosis (CC3) using 3,3′-DAB. Scale bar, 50 μm. (c) Quantification of NQO1, proliferative marker Ki67 and apoptotic marker CC3 in RKO/pNQO1, RKO/shCont and RKO/pshNQO1 xenograft tumours (n=3 each group). n=5 in each tumour. Two-tail t-test. **P<0.01, *P<0.05. #, not significant. All error bars represent the mean±s.e.m. (d) RKO/pshCont1/pshCont2, RKO/pNQO1/pshCont2 and RKO/pNQO1/pshHIF-1 alpha injected subcutaneously into the right flank of athymic, 7-week-old female BALB/C nude mice, and tumour growth was assessed. Tumour volume (TV) was calculated by using the following formula: TV=length × (width)2 × 0.5. Each group contained 10 animals (*P<0.05 with unpaired t-test). (e) Immunohistochemical analyses of RKO/pshCont1/pshCont2, RKO/pNQO1/pshCont2 and RKO/pNQO1/pshHIF-1 alpha xenograft tumours for HIF-1 alpha, proliferation (Ki67), apoptosis (CC3) and vasculature (CD34) using 3,3′-DAB. Arrowheads denote blood vessels. Scale bar, 50 μm. (f) RKO/pNQO1/pshCont or RKO/pNQO1/pshHIF-1 alpha xenograft tumours (n=3–5 each group) were quantified for proliferation (Ki67), apoptosis (CC3) and vascularization (CD34). n=5 in each tumour. Two tailed t-test. **P<0.01, *P<0.05. #, not significant. All error bars represent the mean±s.e.m. (g) Schematic model showing how NQO1 stabilizes HIF-1 alpha in cancer cells. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/27966538), licensed under a CC-BY license. Not internally tested by R&D Systems.

Immunohistochemistry Detection of Mouse EGLN2/PHD1 by Immunohistochemistry View Larger

Detection of Mouse EGLN2/PHD1 by Immunohistochemistry NQO1 promotesin vivo tumour growth.(a) RKO/pNQO1, RKO/shCont and RKO/pshNQO1 cells were injected subcutaneously into the right flank of athymic, 7-week-old female BALB/C nude mice, and tumour growth was assessed. Tumour volume (TV) was calculated by using the following formula: TV=length × (width)2 × 0.5. Each group contained 12 animals. (**P<0.01 with unpaired t-test). (b) Immunohistochemical analyses of RKO/pNQO1, RKO/shCont, and RKO/pshNQO1 xenograft tumours. The sections were stained for NQO1, HIF-1 alpha, proliferation (Ki67) and apoptosis (CC3) using 3,3′-DAB. Scale bar, 50 μm. (c) Quantification of NQO1, proliferative marker Ki67 and apoptotic marker CC3 in RKO/pNQO1, RKO/shCont and RKO/pshNQO1 xenograft tumours (n=3 each group). n=5 in each tumour. Two-tail t-test. **P<0.01, *P<0.05. #, not significant. All error bars represent the mean±s.e.m. (d) RKO/pshCont1/pshCont2, RKO/pNQO1/pshCont2 and RKO/pNQO1/pshHIF-1 alpha injected subcutaneously into the right flank of athymic, 7-week-old female BALB/C nude mice, and tumour growth was assessed. Tumour volume (TV) was calculated by using the following formula: TV=length × (width)2 × 0.5. Each group contained 10 animals (*P<0.05 with unpaired t-test). (e) Immunohistochemical analyses of RKO/pshCont1/pshCont2, RKO/pNQO1/pshCont2 and RKO/pNQO1/pshHIF-1 alpha xenograft tumours for HIF-1 alpha, proliferation (Ki67), apoptosis (CC3) and vasculature (CD34) using 3,3′-DAB. Arrowheads denote blood vessels. Scale bar, 50 μm. (f) RKO/pNQO1/pshCont or RKO/pNQO1/pshHIF-1 alpha xenograft tumours (n=3–5 each group) were quantified for proliferation (Ki67), apoptosis (CC3) and vascularization (CD34). n=5 in each tumour. Two tailed t-test. **P<0.01, *P<0.05. #, not significant. All error bars represent the mean±s.e.m. (g) Schematic model showing how NQO1 stabilizes HIF-1 alpha in cancer cells. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/27966538), licensed under a CC-BY license. Not internally tested by R&D Systems.

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
Sterile PBS to a final concentration of 0.2 mg/mL.
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
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: EGLN2/PHD1

EGLN2 (EGL/EGg Laying-Nine #2; also known as PHD1 and HPH3) is a 43 kDa member of the EglN family of proteins. It is ubiquitously expressed and found principally in the nucleus. EGLN2 hydroxylates proline on HIF-1 alpha. HIF-1 is an alpha / beta heterodimeric transcriptional activator that upregulates genes involved in mitigating the effects of hypoxia. Normally, and in the presence of abundant oxygen, the HIF-1 alpha -chain is hydroxylated by PHD family members, which results in its ubiquitylation and degradation. Under low oxygen tension, EGLN2 activity is decreased, the HIF-1 alpha subunit is retained, and HIF-1 activates genes. Human EGLN2 is 407 amino acids (aa) in length (SwissProt #:Q96KS0). It contains one iron 2-oxoglutarate (Fe2OG) dioxygenase domain (aa 278-376) plus an iron-binding (His297 and His358), and a 2-oxoglutarate-binding (Arg367) site. There is one alternative start site at Met34 that generates a 40 kDa isoform. In addition, there is another potential splice form that shows a 16 aa substitution for aa 1-281. Full-length human EGLN2 shares 91% aa sequence identity with mouse EGLN2.

Long Name
Egl Nine Homolog 2/Prolyl Hydroxylase Domain-containing Protein 1
Entrez Gene IDs
112398 (Human); 112406 (Mouse); 308457 (Rat)
Alternate Names
DKFZp434E026; EC 1.14.11; EC 1.14.11.-; egl nine homolog 2 (C. elegans); egl nine homolog 2; EGLN2; EIT6; Estrogen-induced tag 6; FLJ95603; HIFPH1 EGL nine (C.elegans) homolog 2; HIFPH1; HIF-PH1; HIF-prolyl hydroxylase 1; HPH-1; HPH-3; Hypoxia-inducible factor prolyl hydroxylase 1; PHD1; Prolyl hydroxylase domain-containing protein 1

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Citations for Human EGLN2/PHD1 Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

7 Citations: Showing 1 - 7
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  1. PHD1 regulates p53-mediated colorectal cancer chemoresistance.
    Authors: Deschoemaeker S, Di Conza G, Lilla S et al.
    EMBO Mol Med.
  2. Cystathione ?-synthase regulates HIF-1? stability through persulfidation of PHD2
    Authors: Dey A, Prabhudesai S, Zhang Y et al.
    Science Advances
  3. Cystathione ?-synthase regulates HIF-1? stability through persulfidation of PHD2
    Authors: Dey A, Prabhudesai S, Zhang Y et al.
    Science Advances
  4. Ginsenoside 20(S)-Rg3 suppresses ovarian cancer migration via hypoxia-inducible factor 1 alpha and nuclear factor-kappa B signals
    Authors: T Liu, L Zhao, H Hou, L Ding, W Chen, X Li
    Tumour Biol., 2017-05-01;39(5):1010428317692.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  5. NQO1 inhibits proteasome-mediated degradation of HIF-1?
    Nat Commun, 2016-12-14;7(0):13593.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  6. Docetaxel induced-JNK2/PHD1 signaling pathway increases degradation of HIF-1? and causes cancer cell death under hypoxia
    Authors: Eun-Taex Oh
    Sci Rep, 2016-06-06;6(0):27382.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  7. Ginsenoside 20(S)-Rg3 targets HIF-1alpha to block hypoxia-induced epithelial-mesenchymal transition in ovarian cancer cells.
    Authors: Liu T, Zhao L, Zhang Y, Chen W, Liu D, Hou H, Ding L, Li X
    PLoS ONE, 2014-09-08;9(9):e103887.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot

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