EGLN2/PHD1 Antibody

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.

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 (//pubmed.ncbi.nlm.nih.gov/27966538), licensed under a CC-BY license. Not internally tested by R&D Systems.

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 (//pubmed.ncbi.nlm.nih.gov/27966538), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: WB, IP
• Clonality: Polyclonal
• Host: Sheep
• Conjugate: Unconjugated

EGLN2/PHD1 Antibody Summary

• Immunogen: S. frugiperda insect ovarian cell line Sf 21-derived recombinant human EGLN2/PHD1
  Asp2-Thr407
  Accession # Q96KS0
• Specificity: Detects human EGLN2/PHD1 in direct ELISAs and Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Sheep
• Gene: EGLN2
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunoprecipitation 25 ug/mL
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Reconstitution Instructions: Sterile PBS to a final concentration of 0.2 mg/mL.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for EGLN2/PHD1 Antibody

• 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

Background

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.

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 EGLN2/PHD1 Antibody (AF6394)(7)

Publications using AF6394

• Dey A, Prabhudesai S, Zhang Y et al. Cystathione ?-synthase regulates HIF-1? stability through persulfidation of PHD2 Science Advances 2020-07-03 [PMID: 32937467]
• Dey A, Prabhudesai S, Zhang Y et al. Cystathione ?-synthase regulates HIF-1? stability through persulfidation of PHD2 Science Advances 7/3/2020 [PMID: 32937467]
• T Liu, L Zhao, H Hou, L Ding, W Chen, X Li Ginsenoside 20(S)-Rg3 suppresses ovarian cancer migration via hypoxia-inducible factor 1 alpha and nuclear factor-kappa B signals Tumour Biol., 2017-05-01;39(5):1010428317692. 2017-05-01 [PMID: 28459376] (Western Blot, Human)
• NQO1 inhibits proteasome-mediated degradation of HIF-1? Nat Commun, 2016-12-14;7(0):13593. 2016-12-14 [PMID: 27966538] (Western Blot, Human)
• Eun-Taex Oh Docetaxel induced-JNK2/PHD1 signaling pathway increases degradation of HIF-1? and causes cancer cell death under hypoxia Sci Rep, 2016-06-06;6(0):27382. 2016-06-06 [PMID: 27263528] (Western Blot, Human)
• Deschoemaeker S, Di Conza G, Lilla S et al. PHD1 regulates p53-mediated colorectal cancer chemoresistance. EMBO Mol Med. 8/19/2015 [PMID: 26290450]
• Liu T, Zhao L, Zhang Y, Chen W, Liu D, Hou H, Ding L, Li X Ginsenoside 20(S)-Rg3 targets HIF-1alpha to block hypoxia-induced epithelial-mesenchymal transition in ovarian cancer cells. PLoS ONE, 2014-09-08;9(9):e103887. 2014-09-08 [PMID: 25197976] (Western Blot, Human)

Bioinformatics

• Gene Symbol: EGLN2
• Uniprot:
  • Q96KS0(Human)