HIF-2 alpha/EPAS1 Antibody

Western Blot: HIF-2 alpha/EPAS1 Antibody [NB100-122] - WNT11 is induced by hypoxia or hypoxic mimetics in different cell types. Immunoblot analyses of HeLa cells under normal air or hypoxia for 24 hrs. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep21520) licensed under a CC-BY licence.

Knockout Validated: HIF-2 alpha/EPAS1 Antibody [NB100-122] - HIF-1alpha is the predominant transcriptional regulator of WNT11 expression during hypoxia. EMSCs isolated from the indicated mouse genotypes were infected with lentivirus expressing GFP or Cre recombinase. Non-infected cells and GFP infected cells served as controls. Immunoblot analyses of EMSCs derived from the indicated genotypes treated with 0.1 mM DMOG for 24 hrs. Attenuated WNT11 expression in Hif-1alpha KO EMSCs (lenti-Cre infected Hif-1af/f). Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep21520) licensed under a CC-BY licence.

Dual RNAscope ISH-IHC: HIF-2 alpha/EPAS1 Antibody [NB100-122] - Formalin-fixed paraffin-embedded tissue sections of human placenta were probed for HIF-2 alpha/EPAS1 mRNA (ACD RNAScope Probe, catalog #410598; Fast Red chromogen, ACD catalog # 322750). Adjacent tissue section was processed for immunohistochemistry using Rabbit Polyclonal (Novus Biologicals catalog # NB100-122) at 1:100 dilution with one-hour incubation at room temperature followed by incubation with anti-rabbit IgG VisUCyte HRP Polymer Antibody (Catalog # VC003) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue). Specific staining was localized to trophoblastic cells.

Immunohistochemistry: HIF-2 alpha/EPAS1 Antibody [NB100-122] - Expression of hypoxia-inducible alpha subunits in normal and diseased lung tissue. HIF-2alpha expression are more evident in fibroblasts from idiopathic pulmonary fibrosis (d) than from lung tissue affected by other inflammatory conditions (i.e. chronic bronchitis, panel e) or normal lung tissue (f); as demonstrated by a higher proportion of positive fibroblasts (open arrows) than negative ones (solid black arrows). Image collected and cropped by CiteAb from the following publication (//respiratory-research.biomedcentral.com/articles/10.1186/s12931-019-1100-4) licensed under a CC-BY licence.

Simple Western: HIF-2 alpha/EPAS1 Antibody [NB100-122] - Image shows a specific band for HIF-2 alpha in 0.5 mg/mL of hypoxic HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Immunohistochemistry: HIF-2 alpha/EPAS1 Antibody [NB100-122] - Expression and correlation of 3 antibodies: HIF-1 alpha, HIF-2 alpha and HAF in ccRCC. (A, C & E) Positive nuclear staining to the 3 antibodies: HIF-1 alpha, HIF-2 alpha and HAF, in primary ccRCCs at high magnification (100X). (B, D & F) Heterogenous nuclear staining of 3 antibodies in a tumor at low magnification (20X). (G, H and I). Correlation of 3 antibodies with the Fuhrman grade.Citation: Ambrosetti D, Dufies M, Dadone B, Durand M, Borchiellini D, Amiel J, et al. (2018) The two glycolytic markers GLUT1 and MCT1 correlate with tumor grade and survival in clear-cell renal cell carcinoma. PLoS ONE 13(2): e0193477. //doi.org/10.1371/journal.pone.0193477

Western Blot: HIF-2 alpha/EPAS1 Antibody [NB100-122] - HIF-2 alpha in human retinal and choroidal primary endothelia lysates using.. Image from verified customer review.

Western Blot: HIF-2 alpha/EPAS1 Antibody [NB100-122] - HIF-2 alpha in MDA-MB-231 cell lysate (overexpression and endogenous samples) using. did not react to HIF-1 alpha overexpression. Image from verified customer review.

Western Blot: HIF-2 alpha/EPAS1 Antibody [NB100-122] - Induced WNT11 expression with tumor hypoxia and WNT11 regulates tumor growth. Bevacizumab increased expression of HIF-1alpha and HIF-2alpha and WNT11. First 10 lanes are control tumors, and the last 10 lanes are tumors from bevacizumab-treated animals. Lysates from whole tissue and nuclei are indicated. Alpha -Tubulin, actin and lamin A/C are loading controls. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep21520) licensed under a CC-BY licence.

Chromatin Immunoprecipitation: HIF-2 alpha/EPAS1 Antibody [NB100-122] - MDA-MB-231 cells were exposed to 20% or 1% O2 for 16 hours, and chromatin immunoprecipitation (ChIP) was performed with the indicated antibody (Ab). Primers flanking the HIF binding site were used for qPCR. ChIP image submitted by a verified customer review.

Western Blot: HIF-2 alpha/EPAS1 Antibody [NB100-122] - Lane 1: Cobalt chloride treated COS7 nuclear extracts. Lane 2: Untreated COS7 nuclear extracts.

Western Blot: HIF-2 alpha/EPAS1 Antibody [NB100-122] - 786-O cells without or with VHL overexpression. Image from verified customer review.

Western Blot: HIF-2 alpha/EPAS1 Antibody [NB100-122] - Analysis using the HRP conjugate of NB100-122. Detection of normoxic and hypoxic nuclear rat cell lysates.

Immunohistochemistry-Paraffin: HIF-2 alpha/EPAS1 Antibody [NB100-122] - HIF-2 alpha immunoreactivity in human cardiac myocytes stained with NB100-122.

Knockdown Validated: HIF-2 alpha/EPAS1 Antibody [NB100-122] - Reduction of HIF-2alpha levels leads to protection in UV-triggered apoptosis, but not for apoptosis caused by glucose and serum starvation in 786-O cells. Parental 786-O or those either stably expressing wild-type VHLp19 or stably infected with a control vector (pSuperRetro) or a pool of two HIF-2alpha shRNAs vectors [21] were grown to confluence and lysed. Cell alpha-tubulin.

• Reactivity: Hu, Mu, Rt, Fi, Ha, Pm, Rb, Re, Sh
• Applications: WB, Simple Western, ChIP, ELISA, Flow, GS, IB, ICC/IF, IHC, IHC-Fr, IHC-P, In vitro, IP, PAGE, Dual ISH-IHC, KD, KO
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1.0 mg/ml

HIF-2 alpha/EPAS1 Antibody Summary

• Immunogen: This HIF-2 alpha/EPAS1 Antibody was developed against a peptide derived from the C-terminus of mouse/human HIF-2 alpha protein.
• Specificity: This HIF-2 alpha/EPAS1 Antibody is specific for HIF-2 alpha/EPAS, and does not cross-react with HIF-1 alpha.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: EPAS1
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Western Blot 1 - 2 ug/mL
  • Simple Western 1:50
  • Chromatin Immunoprecipitation 1:10 - 1:500
  • ELISA 1:100 - 1:2000
  • Flow Cytometry
  • Gel Super Shift Assays
  • Immunoblotting
  • Immunocytochemistry/Immunofluorescence 1:100
  • Immunohistochemistry 1:100
  • Immunohistochemistry-Frozen
  • Immunohistochemistry-Paraffin 1:100
  • Immunoprecipitation 5ug/1mg lysate
  • In vitro assay
  • SDS-Page
  • Dual RNAscope ISH-IHC
  • Knockdown Validated
  • Knockout Validated
• Application Notes: Use in Gel Shift Super Assay reported in scientific literature (PMID: 15184875), Use in Immunoblotting reported in scientific literature (PMID 28115701). In WB, this antibody recognizes a band at 118 kDa representing HIF-2 alpha. In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Knock Out Validation was reported in scientific literature (PMID: 26861754). Use in SDS-Page reported in scientific literature (PMID: 27686097). Use In vitro assay reported in scientific literature (PMID: 24998849). Use in Knockdown Validated reported in scientific literature (PMID:31061092).
• Theoretical MW: 96.5 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• Control:

  PC-12 Nuclear Cell Lysate
  COS-7 Nuclear Hypoxic Induced Cell Lysate
  Human Rectum Whole Tissue Lysate (Adult Whole Normal)
  HIF-2 alpha/EPAS1 Knockout HeLa Cell Lysate

Reactivity Notes

Reptile reactivity reported in scientific literature (PMID: 31409194).

Packaging, Storage & Formulations

• Storage: Store at -20 °C.
• Buffer: PBS
• Preservative: 0.05% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for HIF-2 alpha/EPAS1 Antibody

• anti hif 2 alpha
• anti-HIF-2 alpha
• Basic-helix-loop-helix-PAS protein MOP2
• BHLHE73
• Class E basic helix-loop-helix protein 73
• ECYT4
• endothelial PAS domain protein 1
• endothelial PAS domain-containing protein 1
• EPAS1
• EPAS-1
• HIF 2A
• HIF-1-alpha-like factor
• HIF-1alpha-like factor
• HIF2 alpha
• HIF-2 alpha
• hif2a angiogenesis
• HIF2A
• HIF-2-alpha
• HIF2-alpha
• HLF
• hypoxia-inducible factor 2 alpha
• Hypoxia-inducible factor 2-alpha
• Member of PAS protein 2
• MOP2
• PAS domain-containing protein 2
• PASD2

Background

Hypoxia contributes to the pathophysiology of human disease, including myocardial and cerebral ischemia, cancer, pulmonary hypertension, congenital heart disease and chronic obstructive pulmonary disease (1). In cancer, and particularly solid tumors, hypoxia plays a critical role in the regulation of genes involved in stem cell renewal, epithelial to mesenchymal transition (EMT), metastasis and angiogenesis. In the tumor microenvironment (TME), hypoxia influences the properties and function of stromal cells (e.g., fibroblasts, endothelial and immune cells) and is a strong determinant of tumor progression (2,3).

HIF-1 or hypoxia inducible factor 1, is a transcription factor commonly referred to as a "master regulator of the hypoxic response" for its central role in the regulation of cellular adaptations to hypoxia. Similarly, HIF-2 alpha plays a role in cellular responses to hypoxia, but whereas HIF-1 alpha is ubiquitously expressed, HIF-2 alpha is predominantly expressed in the vascular endothelium at embryonic stages and after birth in select cells and tissue types (e.g., fibroblasts, hepatocytes and myocytes at 96kDa) (4). Following a similar mechanism to HIF-1 alpha, HIF-2 alpha is stabilized under hypoxic conditions by the formation of a heterodimer with an ARNT/HIF-1 beta subunit. Stable HIF-2 alpha-ARNT/HIF-1 beta heterodimers engage p300/CBP in the nucleus for binding to hypoxic response elements (HREs), inducing transcription, and thus regulation of genes (e.g., EPO, VEGFA). HIF-1 predominantly transactivates genes involved in glycolytic control and pro-apoptotic genes (e.g., LDHA and BNIP3), and HIF-2 regulates the expression of genes involved in invasion and stemness (e.g., MMP2, and OCT4). Common gene targets for HIF-1 and HIF-2 include VEGFA and GLUT1 (5).

The HIF-2 alpha subunit is rapidly targeted and degraded by the ubiquitin proteasome system under normoxic conditions. This process is mediated by oxygen-sensing enzymes, prolyl hydroxylase domain enzymes (PHDs), which catalyze the hydroxylation of key proline residues (Pro-405 and Pro-531) within the oxygen-dependent degradation domain of HIF-2 alpha (5). Once hydroxylated, HIF-2 alpha binds the von Hippel-Lindau tumor suppressor protein (pVHL) for subsequent ubiquitination and proteasomal degradation (5,6).

References

1. Semenza, G. L., Agani, F., Feldser, D., Iyer, N., Kotch, L., Laughner, E., & Yu, A. (2000). Hypoxia, HIF-1, and the pathophysiology of common human diseases. Advances in Experimental Medicine and Biology.

2.Muz, B., de la Puente, P., Azab, F., & Azab, A. K. (2015). The role of hypoxia in cancer progression, angiogenesis, metastasis, and resistance to therapy. Hypoxia. https://doi.org/10.2147/hp.s93413

3. Huang, Y., Lin, D., & Taniguchi, C. M. (2017). Hypoxia inducible factor (HIF) in the tumor microenvironment: friend or foe? Science China Life Sciences. https://doi.org/10.1007/s11427-017-9178-y

4. Hu, C.-J., Wang, L.-Y., Chodosh, L. A., Keith, B., & Simon, M. C. (2003). Differential Roles of Hypoxia-Inducible Factor 1 (HIF-1) and HIF-2 in Hypoxic Gene Regulation. Molecular and Cellular Biology. https://doi.org/10.1128/mcb.23.24.9361-9374.2003

5. Koh, M. Y., & Powis, G. (2012). Passing the baton: The HIF switch. Trends in Biochemical Sciences. https://doi.org/10.1016/j.tibs.2012.06.004

6. Koyasu, S., Kobayashi, M., Goto, Y., Hiraoka, M., & Harada, H. (2018). Regulatory mechanisms of hypoxia-inducible factor 1 activity: Two decades of knowledge. Cancer Science. https://doi.org/10.1111/cas.13483

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 HIF-2 alpha/EPAS1 Antibody (NB100-122)(626)

Publications using NB100-122

• Acosta-Iborra B, Tiana M, Maeso-Alonso L et al. Hypoxia compensates cell cycle arrest with progenitor differentiation during angiogenesis FASEB J. Mar 29 2020 [PMID: 32223018]
• Chen L, Wang Z, Xu Q et Al. The failure of DAC to induce OCT2 expression and its remission by hemoglobin-based nanocarriers under hypoxia in renal cell carcinoma Theranostics Feb 18 2020 [PMID: 32206108] (WB, Mouse)
• Fabbri L, Dufies M, Lacas-Gervais S et Al. Identification of a new aggressive axis driven by ciliogenesis and absence of VDAC1-deltaC in clear cell Renal Cell Carcinoma patients Theranostics Feb 3 2020 [PMID: 32194829] (WB, ICC/IF, Human)
• Park Jh, Kim Mj, Kim Wj et Al. Isolinderalactone suppresses human glioblastoma growth and angiogenic activity in 3D microfluidic chip and in vivo mouse models Cancer Lett. Mar 12 2020 [PMID: 32173479] (WB, Human)
• Sun X, Sun Bl, Babicheva A et Al. Direct eNAMPT Involvement in Pulmonary Hypertension and Vascular Remodeling: Transcriptional Regulation by SOX and HIF2 alpha Am. J. Respir. Cell Mol. Biol. Mar 6 2020 [PMID: 32142369]
• Gojkovic M, Darmasaputra Gs, Velica P et Al. Deregulated hypoxic response in myeloid cells: A model for high-altitude pulmonary oedema (HAPE) Acta Physiol (Oxf) Mar 4 2020 [PMID: 32129933] (WB, ICC/IF, Mouse)
• Sadaghianloo N, Contenti J, Dufies M et al. Co-culture of human fibroblasts, smooth muscle and endothelial cells promotes osteopontin induction in hypoxia J. Cell. Mol. Med. Feb 7 2020 [PMID: 32032472] (WB, Human)
• Roelants C, Pillet C, Franquet Q et al. Ex-Vivo Treatment of Tumor Tissue Slices as a Predictive Preclinical Method to Evaluate Targeted Therapies for Patients with Renal Carcinoma Cancers (Basel) Jan 17 2020 [PMID: 31963500] (IHC, Human)
• Sivaraj KK, Dharmalingam B, Mohanakrishnan V et al. YAP1 and TAZ negatively control bone angiogenesis by limiting hypoxia-inducible factor signaling in endothelial cells Elife Jan 20 2020 [PMID: 31958058] (ICC/IF, Mouse)
• Brown E, Rowan C, Strowitzki MJ et al. Mucosal inflammation downregulates PHD1 expression promoting a barrier-protective HIF-1 alpha response in ulcerative colitis patients FASEB J. Jan 16 2020 [PMID: 31944416] (WB, Human)
• Senavirathna LK, Huang C, Pushparaj S et al. Hypoxia and transforming growth factor beta 1 regulation of long non-coding RNA transcriptomes in human pulmonary fibroblasts Physiol Rep Jan 1 2020 [PMID: 31925944] (WB, Human)
• Jeon GW, Sheldon RA, Ferriero DM Hypoxia-inducible factor: role in cell survival in superoxide dismutase overexpressing mice after neonatal hypoxia-ischemia Korean J Pediatr Dec 1 2019 [PMID: 31870086] (WB, Mouse)
• Yoo HC, Park SJ, Nam M et al. A Variant of SLC1A5 Is a Mitochondrial Glutamine Transporter for Metabolic Reprogramming in Cancer Cells Cell Metab. Dec 13 2019 [PMID: 31866442] (ChIP, Human)
• Hart PC, Kenny HA, Grassl N, et al. Mesothelial Cell HIF1 alpha Expression Is Metabolically Downregulated by Metformin to Prevent Oncogenic Tumor-Stromal Crosstalk Cell Rep Dec 17 2019 [PMID: 31851935] (WB, Human)
• Varghese J, Varghese James J, Karthikeyan M et al. Iron homeostasis is dysregulated, but the iron-hepcidin axis is functional, in chronic liver disease Journal of Trace Elements in Medicine and Biology Nov 1 2019 [PMID: 31835128] (WB, Human)
• Cannito S, Foglia B, Villano G et al. SerpinB3 Differently Up-Regulates Hypoxia Inducible Factors -1 alpha and -2 alpha in Hepatocellular Carcinoma: Mechanisms Revealing Novel Potential Therapeutic Targets Cancers (Basel) Dec 4 2019 [PMID: 31817100] (WB, Human)
• Morikawa M, Mitani Y, Holmborn K et al. The ALK-1/SMAD/ATOH8 axis attenuates hypoxic responses and protects against the development of pulmonary arterial hypertension Sci Signal Nov 12 2019 [PMID: 31719172]
• Tashiro N, Segawa R, Tobita R et al. Hypoxia inhibits TNF-alpha-induced TSLP expression in keratinocytes PLoS ONE Nov 4 2019 [PMID: 31682627] (WB, Human, Mouse)
• Zhang X, Zhang Y, Wang P et al. Adipocyte Hypoxia-Inducible Factor 2 alpha Suppresses Atherosclerosis by Promoting Adipose Ceramide Catabolism Cell Metab. Oct 23 2019 [PMID: 31668872]
• Han S, Huang T, Hou F et al. The prognostic value of hypoxia-inducible factor-1 alpha in advanced cancer survivors: a meta-analysis with trial sequential analysis Ther Adv Med Oncol 2019 Sep 24 [PMID: 31579115] (IHC-P, Human)
• Gao RY, Wang M, Liu Q et al. Hypoxia-Inducible Factor (HIF)-2 alpha Reprograms Liver Macrophages to Protect Against Acute Liver Injury via the Production of Interleukin-6 Hepatology Sep 17 2019 [PMID: 31529728] (WB)
• Seo JB, Riopel M, Cabrales P et al. Knockdown of Ant2 Reduces Adipocyte Hypoxia And Improves Insulin Resistance in Obesity Nat Metab [PMID: 31528845] (WB, Human)
• David BT, Curtin JJ, Goldberg DC et al. Hypoxia Inducible Factor 1 alpha (HIF-1 alpha) counteracts the acute death of cells transplanted into the injured spinal cord eNeuro [PMID: 31488552] (WB)
• Schwartz AJ, Converso-Baran K, Michele DE, Shah YM A genetic mouse model of severe iron-deficiency anemia reveals tissue-specific transcriptional stress responses and cardiac remodeling J. Biol. Chem. Aug 15 2019 [PMID: 31416832] (IHC-Fr, Mouse)
• Novianti T, Juniantito V, Jusuf AA et al. Expression and role of HIF-1alpha and HIF-2alpha in tissue regeneration: a study of hypoxia in house gecko tail regeneration Organogenesis Aug 13 2019 [PMID: 31409194] (WB, IHC-P, Reptile)
  
  Details:
  The role of HIF-1 alpha and HIF-2 alpha in overcoming hypoxia during tail autotomy in the Hemidactylus platyurus is explored.
• Qi D, Wei M, Jiao S et al. Hypoxia inducible factor 1 alpha in vascular smooth muscle cells promotes angiotensin II-induced vascular remodeling via activation of CCL7-mediated macrophage recruitment Cell Death Dis Jul 18 2019 [PMID: 31320613] (ChIP, Mouse)
• Lee YS, Riopel M, Cabrales P, Bandyopadhyay GK, Hepatocyte-specific HIF-1 alpha ablation improves obesity-induced glucose intolerance by reducing first-pass GLP-1 degradation Sci Adv Jul 1 2019 [PMID: 31281892] (WB, Mouse)
• Aquino-Galvez A, Gonzalez-Avila G, Jimenez-Sanchez LL et al. Dysregulated expression of hypoxia-inducible factors augments myofibroblasts differentiation in idiopathic pulmonary fibrosis Respir. Res. Jun 24 2019 [PMID: 31234835] (WB, Human)
• Maugeri G, D'Amico AG, Federico C et al. Involvement of A3 Adenosine Receptor in Neuroblastoma Progression via Modulation of the Hypoxic/Angiogenic Pathway J. Mol. Neurosci. Jun 5 2019 [PMID: 31166001] (WB, Human)
• Yamamoto A, Hester J, Macklin PS, et al. Systemic silencing of PHD2 causes reversible immune regulatory dysfunction J. Clin. Invest. 2019 Jun 04 [PMID: 31162141] (WB, Mouse)
• Fang C, Manes TD, Liu L et al. A RASSF1A-HIF1 alpha loop drives Warburg effect in cancer and pulmonary hypertension Nat Commun May 13 2019 [PMID: 31086178] (WB, Human)
• Liu M, Zhang L, Marsboom G, Jambusaria A et al. Sox17 is required for endothelial regeneration following inflammation-induced vascular injury Nat Commun May 9 2019 [PMID: 31073164] (WB, Mouse)
• Muenchau S, Deutsch R, de Castro IJ et al. Hypoxic Environment Promotes Barrier Formation in Human Intestinal Epithelial Cells through Regulation of MicroRNA 320a Expression Mol. Cell. Biol. Jul 15 2019 [PMID: 31061092] (WB, KD, Human)
• Fujimoto TN, Colbert LE, Huang Y et al. Selective EGLN Inhibition Enables Ablative Radiotherapy and Improves Survival in Unresectable Pancreatic Cancer Cancer Res. May 1 2019 [PMID: 31043430] (WB, Mouse)
• Bechmann N, Poser I, Seifert V et al. Impact of Extrinsic and Intrinsic Hypoxia on Catecholamine Biosynthesis in Absence or Presence of Hif2 alpha in Pheochromocytoma Cells Cancers (Basel) Apr 28 2019 [PMID: 31035382] (IHC-P, WB, Mouse)
• Chaurasia, SN;Kushwaha, G;Kulkarni, PP;Mallick, RL;Latheef, NA;Mishra, JK;Dash, D; Platelet HIF-2 alpha promotes thrombogenicity through PAI-1 synthesis and extracellular vesicle release Haematologica Apr 19 2019 [PMID: 31004026] (WB, Human)
• Sumi Chisato, Matsuo Yoshiyuki, Kusunoki Munenori et al. Cancerous phenotypes associated with hypoxia-inducible factors are not influenced by the volatile anesthetic isoflurane in renal cell carcinoma. PloS One 2019 Apr 15 [PMID: 30986231] (Human)
• Gkotinakou, IM;Befani, C;Simos, G;Liakos, P; ERK1/2 phosphorylates HIF-2 alpha and regulates its activity by controlling its CRM1-dependent nuclear shuttling J. Cell. Sci. Apr 8 2019 [PMID: 30962349] (WB, Human)
• Chang Song Ho, Mori Daisuke, Kobayashi Hiroshi et al. Excessive mechanical loading promotes osteoarthritis through the gremlin-1-NF- kappa B pathway. Frontiers in Immunology 2019 Mar 29 [PMID: 30926814] (IHC-P, Mouse)
• Chachami, G;Stankovic-Valentin, N;Karagiota, A;Basagianni, A;Plessmann, U;Urlaub, H;Melchior, F;Simos, G; Hypoxia-induced changes in SUMO conjugation affect transcriptional regulation under low oxygen Mol. Cell Proteomics Mar 29 2019 [PMID: 30926672] (WB, Human)
• Arthur SA, Blaydes JP, Houghton FD. Glycolysis Regulates Human Embryonic Stem Cell Self-Renewal under Hypoxia through HIF-2 alpha and the Glycolytic Sensors CTBPs Stem Cell Reports Mar 1 2019 [PMID: 30880076] (WB, Human)
• Natarajan S, Foreman KM, Soriano MI et al. Collagen remodeling in the hypoxic tumor-mesothelial niche promotes ovarian cancer metastasis Cancer Res. Mar 12 2019 [PMID: 30862717] (IHC, Human)
• Syafruddin SE, Rodrigues P, Vojtasova E et al. A KLF6-driven transcriptional network links lipid homeostasis and tumour growth in renal carcinoma Nat Commun Mar 11 2019 [PMID: 30858363] (ChIP, Human)
• Halvarsson C, Rorby E, Eliasson P et al. Putative role of NF-kappa B but not HIF-1alpha in hypoxia-dependent regulation of oxidative stress in hematopoietic stem and progenitor cells Antioxid. Redox Signal. Mar 2 2019 [PMID: 30827134] (WB, Mouse)
• Ndiaye PD, Dufies M, Giuliano S et al. VEGFC acts as a double-edged sword in renal cell carcinoma aggressiveness Theranostics Jan 21 2019 [PMID: 30809300] (WB, Human)
• Laitakari A, Ollonen T, Kietzmann T et al. Systemic inactivation of hypoxia-inducible factor prolyl 4-hydroxylase 2 in mice protects from alcohol-induced fatty liver disease Redox Biol Feb 19 2019 [PMID: 30802717] (WB, Mouse)
• Xiong Y, Liu L, Xia Y et al. Tumor infiltrating mast cells determine oncogenic HIF-2a-conferred immune evasion in clear cell renal cell carcinoma Cancer Immunol. Immunother. Feb 13 2019 [PMID: 30758643] (IHC-P, Human)
• Hong B, Zhou J, Ma K et al. TRIB3 Promotes the Proliferation and Invasion of Renal Cell Carcinoma Cells via Activating MAPK Signaling Pathway. Int. J. Biol. Sci. Jan 8 2019 [PMID: 30745845] (WB, Human)
• Lee BJ, Jun HO, Kim JH, Kim JH. Astrocytic cystine/glutamate antiporter is a key regulator of erythropoietin expression in the ischemic retina FASEB J. Feb 11 2019 [PMID: 30742774] (WB, Human)
• Tian Z, Yao L, Shen Y et al. Histone H3K9 demethylase JMJD1A is a co-activator of erythropoietin expression under hypoxia Int. J. Biochem. Cell Biol. Apr 1 2019 [PMID: 30716474] (WB, Human)
• Jarman EJ, Ward C, Turnbull AK et al. HER2 regulates HIF-2a and drives an increased hypoxic response in breast cancer. Breast Cancer Res. Jan 22 2019 [PMID: 30670058] (WB, Human)
• Ma X, Das NK, Castillo C et al. SMAD family member 3 (SMAD3) and SMAD4 repress HIF2a-dependent iron-regulatory genes. J. Biol. Chem. Jan 18 2019 [PMID: 30659096] (WB, Human)
• Guo H, Ci X, Ahmed M et al. ONECUT2 is a driver of neuroendocrine prostate cancer. Nat Commun Jan 17 2019 [PMID: 30655535] (ChIP, Human)
• Santana-Codina N, Gableske S, Quiles Del Rey M et al. NCOA4 maintains murine erythropoiesis via cell autonomous and non-autonomous mechanisms. Haematologica Jan 10 2019 [PMID: 30630985] (WB, Mouse)
• Olcina MM, Balanis NG, Kim RK et al. Mutations in an Innate Immunity Pathway Are Associated with Poor Overall Survival Outcomes and Hypoxic Signaling in Cancer Cell Rep Dec 26 2018 [PMID: 30590044] (WB, Human)
• Karagiota A, Kourti M, Simos G, Mylonis I. HIF-1a-derived cell-penetrating peptides inhibit ERK-dependent activation of HIF-1 and trigger apoptosis of cancer cells under hypoxia Cell. Mol. Life Sci. Dec 10 2018 [PMID: 30535970] (WB, Human)
• Choi SA, Kim YH, Park YH et al. Novel crosstalk between Vps26a and Nox4 signaling during neurogenesis. Cell Death Differ. Nov 21 2018 [PMID: 30464227] (ICC/IF, Mouse)
• Murahashi Y, Yano F, Kobayashi H et al. Intra-articular administration of IkBa kinase inhibitor suppresses mouse knee osteoarthritis via downregulation of the NF-kB/HIF-2a axis. Sci Rep. Nov 7 2018 [PMID: 30405206] (IHC-P, Mouse)
• Deygas M, Gadet R, Gillet G et al. Redox regulation of EGFR steers migration of hypoxic mammary cells towards oxygen. Nat Commun. Oct 31 2018 [PMID: 30382089] (WB, Human)
• Liao L, Liu ZZ, Langbein L et al. Multiple tumor suppressors regulate a HIF-dependent negative feedback loop via ISGF3 in human clear cell renal cancer. Elife. Oct 25 2018 [PMID: 30355451] (WB, Human)
• Schwartz AJ, Das Nk, Ramakrishnan Sk et al. Hepatic hepcidin/intestinal HIF-2a axis maintains iron absorption during iron deficiency and overload. J. Clin. Invest. Oct 23 2018 [PMID: 30352047] (Mouse)
• Carolina E, kato T, khanh VC et al. Glucocorticoid Impaired the Wound Healing Ability of Endothelial Progenitor Cells by Reducing the Expression of CXCR4 in the PGE2 Pathway. Front Med (Lausanne). Sep 28 2018 [PMID: 30324106] (WB, Human)
• Fortenbery GW, Sarathy B, Carraway kR, Mansfield kD. Hypoxic stabilization of mRNA is HIF-independent but requires mtROS. Cell. Mol. Biol. Lett. Oct 4 2018 [PMID: 30305827] (WB, Human)
• Sethuraman A, Brown M, krutilina R et al. BHLHE40 confers a pro-survival and pro-metastatic phenotype to breast cancer cells by modulating HBEGF secretion. Breast Cancer Res. Oct 1 2018 [PMID: 30285805] (WB, Human)
• Xu M, Tan X, Li N et al. Differential regulation of estrogen in iron metabolism in astrocytes and neurons J. Cell. Physiol. Aug 21 2018 [PMID: 30132882] (WB, Rat)
• Zaini MN, Patel SA, Syafruddin SE et al. Endogenous HIF2A reporter systems for high-throughput functional screening Sci Rep Aug 13 2018 [PMID: 30104738] (WB, Human)
• Anea C, Merloiu A, Fulton D et al. Immunohistochemistry of the circadian clock in mouse and human vascular tissues. VP Jul 20 2018 [PMID: 30101218] (IHC, Mouse)
• Regan Anderson TM, Ma S, Perez kerkvliet C et al. Taxol Induces Brk-dependent Prosurvival Phenotypes in TNBC Cells through an AhR/GR/HIF-driven Signaling Axis. Mol. Cancer Res. Jul 10 2018 [PMID: 29991529] (WB, Human)
• Miess H, Dankworth B, Gouw AM et al. The glutathione redox system is essential to prevent ferroptosis caused by impaired lipid metabolism in clear cell renal cell carcinoma Oncogene Jun 5 2018 [PMID: 29872221] (WB, Human)
• Kumar VP, Biswas S, Sharma NK et al. PEGylated IL-11 (BBT-059): A Novel Radiation Countermeasure for Hematopoietic Acute Radiation Syndrome Health Phys Jul 1 2018 [PMID: 29787432] (WB, Mouse)
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  Details:
  HIF-2 alpha/EPAS1 was used at 1:500 dilution for WB assays in experiments involving human kidney cell lines namely ACHN, Caki-1 and HEK293T.
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• Cavadas MA, Mesnieres M, Crifo B et al. REST mediates resolution of HIF-dependent gene expression in prolonged hypoxia. Sci Rep. 2015 Dec 09 [PMID: 26647819] (WB, Human)
• Vukovic M, Guitart AV, Sepulveda C et al. Hif-1alpha and Hif-2alpha synergize to suppress AML development but are dispensable for disease maintenance. J. Exp. Med. 2015 Dec 14 [PMID: 26642852] (WB, Human)
• Bhanu NV, Sidoli S, Garcia BA. Histone modification profiling reveals differential signatures associated with human embryonic stem cell self-renewal and differentiation. Proteomics. 2016 Feb 01 [PMID: 26631989]
• Sun W, Kosyna FK, Jelkmann W, Depping R. Prolyl-4-Hydroxylase 2 Potentially Contributes to Hepatocellular Carcinoma-Associated Erythrocytosis by Maintaining Hepatocyte Nuclear Factor-4alpha Expression. Cell. Physiol. Biochem. 2015 Dec 24 [PMID: 26624507] (WB, ChIP, Human)
• Mandl M, Lieberum M, Dunst J, Depping R. The expression level of the transcription factor Aryl hydrocarbon receptor nuclear translocator (ARNT) determines cellular survival after radiation treatment. Radiat Oncol. 2015 Nov 17 [PMID: 26572229] (WB, Human)
• Cheng S, Xing W, Pourteymoor S et al. Conditional Deletion of Prolyl Hydroxylase Domain-containing Protein 2 (Phd2) Gene Reveals its Essential Role in Chondrocyte Function and Endochondral Bone Formation. Endocrinology. 2015 Nov 12 [PMID: 26562260] (WB, Mouse)
• Aquino-Galvez A, Gonzalez-avila G. Effects of 2-methoxyestradiol on apoptosis and HIF-1a and HIF-2a expression in lung cancer cells under normoxia and hypoxia. ONCOLOGY REPORTS. 2015 Aug 20 [PMID: 26548300] (WB, ICC/IF, Human)
• Highet AR, Khoda SM, Buckberry S et al. Hypoxia induced HIF-1/HIF-2 activity alters trophoblast transcriptional regulation and promotes invasion. Eur. J. Cell Biol. 2015 Dec 01 [PMID: 26531845] (IHC-P, Human)
  
  Details:
  HIF2 alpha antibody was used for IHC-P on early / late first trimester and term uncomplicated control placentas. The IHC assay involved antigen retrieval through boiling the slides with citrate buffer pH 6.5 in microwave, blocking of endogenous peroxidase using 6% H2O2 for 1 hour, use of primary at 1:1500 dilution, and detection using biotinylated secondary, Streptavidin-HRP conjugate-DAB followed by counterstaining with haematoxylin (Fig. 5).
• Huh Yun Hyun, Lee Gyuseok, Lee Keun-Bae et al. HIF-2a-induced chemokines stimulate motility of fibroblast-like synoviocytes and chondrocytes into the cartilage-pannus interface in experimental rheumatoid arthritis mouse models. Arthritis Research & Therapy 2015 [PMID: 26510617] (IHC, Mouse)
• Zhang J, Wang C, Chen X et al. EglN2 associates with the NRF1-PGC1alpha complex and controls mitochondrial function in breast cancer. EMBO J 2015 Dec 02 [PMID: 26492917] (WB, Human)
• Saini Y, Proper SP, Dornbos P et al. Loss of Hif-2alpha Rescues the Hif-1alpha Deletion Phenotype of Neonatal Respiratory Distress In Mice. PLoS One 2015 [PMID: 26422241] (IHC, Mouse)
• Stengel C, Newman SP, Leese MP et al. The in vitro and in vivo activity of the microtubule disruptor STX140 is mediated by Hif-1 alpha and CAIX expression Anticancer Res. 2016 Mar 31 [PMID: 26408684] (WB, Human)
• Frey JL, Stonko DP, Faugere MC, Riddle RC Hypoxia-inducible factor-1alpha restricts the anabolic actions of parathyroid hormone. Bone Res 2014 May 13 [PMID: 26273518] (IP)
• Lu H Samanta D Xiang L et al. Chemotherapy triggers HIF-1-dependent glutathione synthesis and copper chelation that induces the breast cancer stem cell phenotype. Proc Natl Acad Sci U S A. 2015 Jul 30 [PMID: 26229077] (WB, Human)
• Hogel H, Miikkulainen P, Bino L, Jaakkola PM. Hypoxia inducible prolyl hydroxylase PHD3 maintains carcinoma cell growth by decreasing the stability of p27. Mol. Cancer. 2015 Jul 30 [PMID: 26223520] (WB, Human)
• Ramakrishnan SK, Anderson ER, Martin A et al. Maternal intestinal HIF-2a is necessary for sensing iron demands of lactation in mice Proc. Natl. Acad. Sci. U.S.A. 2015 Jul 14 [PMID: 26124130] (IHC-Fr, Mouse)
• Zhang X, Zhu J, Liu F et al. Reduced EGFR signaling enhances cartilage destruction in a mouse osteoarthritis model. Bone Res 2014 Aug 05 [PMID: 26120493] (IHC)
• Otto T, Gellhaus A, Luschen N et al. Oxygen Sensitivity of Placental Trophoblast Connexins 43 and 46: A Role in Preeclampsia? J Cell Biochem 2015 May 27 [PMID: 26018820]
• Matsuyama T, Tanaka T, Tatsumi K et al. Midazolam inhibits the hypoxia-induced up-regulation of erythropoietin in the central nervous system. Eur. J. Pharmacol. 2015 May 19 [PMID: 26001375] (WB, Mouse)
• Lin AE, Beasley FC, Olson J et al. Role of Hypoxia Inducible Factor-1alpha (HIF-1alpha) in Innate Defense against Uropathogenic Escherichia coli Infection. PLoS Pathog. 2015 Apr 01 [PMID: 25927232] (WB, Human)
• Forristal CE, Brown AL, Helwani FM et al. Hypoxia inducible factor (HIF)-2alpha accelerates disease progression in mouse models of leukemia and lymphoma but is not a poor prognosis factor in human AML. Leukemia. 2015 Apr 29 [PMID: 25921247] (WB, Mouse)
• Koizume S, Ito S, Nakamura Y et al. Lipid starvation and hypoxia synergistically activate ICAM1 and multiple genes in an Sp1-dependent manner to promote the growth of ovarian cancer Mol. Cancer 2015 Apr 17 [PMID: 25879517] (IHC-P, ICC/IF, Human)
• Shao J, Zhang Y, Yang T et al. HIF-1alpha disturbs osteoblasts and osteoclasts coupling in bone remodeling by up-regulating OPG expression In Vitro Cell. Dev. Biol. Anim. 2015 Apr 10 [PMID: 25859979] (WB, Mouse)
• Rad E, Dodd K, Thomas L et al. STAT3 and HIF1alpha Signaling Drives Oncogenic Cellular Phenotypes in Malignant Peripheral Nerve Sheath Tumors. Mol Cancer Res 2015 Jul [PMID: 25833823] (Human)
• Wu S, Zhang J, Huang C et al. Prolyl hydroxylase domain-2 silencing induced by hydrodynamic limb vein injection enhances vascular regeneration in critical limb ischemia mice through activation of multiple genes. Curr Gene Ther 2015 [PMID: 25832622] (Mouse)
• McMurray F, Demetriades M, Aik W et al. Pharmacological Inhibition of FTO. PLoS One. 2015 Apr 01 [PMID: 25830347] (WB, Mouse)
• Qiu B, Ackerman D, Sanchez DJ et al. HIF-2alpha dependent lipid storage promotes endoplasmic reticulum homeostasis in clear cell renal cell carcinoma Cancer Discov 2015 [PMID: 25829424] (WB, Human)
• Bonora M, Wieckowsk MR, Chinopoulos C et al. Molecular mechanisms of cell death: central implication of ATP synthase in mitochondrial permeability transition. Oncogene 2015 Mar 19 [PMID: 25790189]
• Zeng H, Vaka VR, He X et al. High-fat diet induces cardiac remodelling and dysfunction: assessment of the role played by SIRT3 loss. J. Cell. Mol. Med. 2015 Mar 17 [PMID: 25782072] (WB, Mouse)
• Cotroneo E, Ashek A, Wang L et al. Iron Homeostasis and Pulmonary Hypertension: Iron Deficiency Leads to Pulmonary Vascular Remodeling in the Rat. Circ. Res. 2015 Mar 12 [PMID: 25767292] (WB, Rat)
• Doonachar A, Gallo MD, Doukas D et al. Differential effects of HIF-alpha isoforms on apoptosis in renal carcinoma cell lines Cancer Cell Int. 2015 Mar 02 [PMID: 25729330] (WB, Human)
• Jimenez K, Khare V, Evstatiev R et al. Increased expression of HIF2alpha during iron deficiency-associated megakaryocytic differentiation J. Thromb. Haemost. 2015 Feb 25 [PMID: 25715026] (IHC-P, Rat)
• Matak P, Heinis M, Mathieu JR et al. Myeloid HIF-1 Is Protective in Helicobacter pylori-Mediated Gastritis J. Immunol. 2015 Feb 20 [PMID: 25710915] (IHC-P, Human)
• Chen Rui, Xu Min, Nagati Jason S et al. The acetate/ACSS2 switch regulates HIF-2 stress signaling in the tumor cell microenvironment. PLoS One. 2014 [PMID: 25689462]
• Nozawa-Suzuki N, Nagasawa H, Ohnishi K, Morishige KI. The inhibitory effect of hypoxic cytotoxin on the expansion of cancer stem cells in ovarian cancer Biochem. Biophys. Res. Commun. 2015 Jan 22 [PMID: 25619133] (WB, Human)
• Neelam S, Brooks MM, Cammarata PR. Lenticular cytoprotection, part 2: Link between glycogen synthase kinase-3b, epithelial to mesenchymal transition, and mitochondrial depolarization. Molecular Vision. 2014-Nov-31 [PMID: 25593505] (WB, Human)
  
  Details:
  HIF-2 alpha/EPAS1 antibody used for WB on lysates of HLE-B3 cells, a human lens epithelial cell line immortalized by the SV-40 virus, treated or not with HIF-2 alpha translation inhibitor (CAS882268-69-1) at 0.5, 5 and 50 uM concentrations (Figure 11A)
• Xiang L, Gilkes DM, Hu H et al. Hypoxia-inducible factor 1 mediates TAZ expression and nuclear localization to induce the breast cancer stem cell phenotype. Oncotarget. 2014 [PMID: 25587023] (ChIP, IP, WB, Human)
• Gong H, Rehman J, Tang H et al. HIF2alpha signaling inhibits adherens junctional disruption in acute lung injury. J Clin Invest 2015 Feb [PMID: 25574837]
• Yoo S, Takikawa S, Geraghty P et al. Integrative Analysis of DNA Methylation and Gene Expression Data Identifies EPAS1 as a Key Regulator of COPD PLoS Genet. 2015 Jan 01 [PMID: 25569234] (WB, Human)
  
  Details:
  HIF-2 alpha/EPAS1 antibody used for IHC-P on human lung tissues from Chronic Obstructive Pulmonary Disease/COPD and non-COPD patients - antigen retrieval with 10 mM citrate buffer, endogenous peroxidase blocked with 0.3% H2O2-methanol, blocked with 5% goat serum diluted in PBS containing 0.1% Tween-20 for 30 minutes, primary antibody used at 1:100 dilution with 1 hour RT incubation, detection with anti-rabbit-HRP secondary and Vectastain ABC Elite Kit. HIF-2 alpha/EPAS1 protein levels were found to be lower in the lung tissues of COPD patients compared to control subjects.
• Tsuboi I, Yamashita T, Nagano M et al. Impaired expression of HIF -2alpha induces compensatory expression of HIF-1alpha for the recovery from anemia J. Cell. Physiol. 2015 Jan 03 [PMID: 25557133] (WB, ChIP, Mouse)
• Cannito S, Turato C, Paternostro C et al. Hypoxia up-regulates SERPINB3 through HIF-2alpha in human liver cancer cells. Oncotarget. 2015 Feb 10 [PMID: 25544768] (ChIP, Human)
• Singleton DC, Rouhi P, Zois CE et al. Hypoxic regulation of RIOK3 is a major mechanism for cancer cell invasion and metastasis. Oncogene. 2014 Dec 08 [PMID: 25486436] (WB, Human)
  
  Details:
  HIF-2 alpha/EPAS1 antibody used for WB on MDA-MB-231 cells subjected to hypoxia. The antibody also used on MDA-MB-231 cells expressing siRNA targeting HIF1 alpha or HIF1 alpha and HIF2 alpha (Figure 1)
• Samanta D, Gilkes D, Chaturvedi P, Xiang L, Semenza G. Hypoxia-inducible factors are required for chemotherapy resistance of breast cancer stem cells. Proc Natl Acad Sci U S A. 2014 Dec 1 [PMID: 25453096] (WB, Human)
• Koh MY, Nguyen V, Lemos R et al. Hypoxia-Induced SUMOylation of E3 Ligase HAF Determines Specific Activation of HIF2 in Clear-Cell Renal Cell Carcinoma. Cancer Res. 2014 Nov 24 [PMID: 25421578] (WB, Human)
  
  Details:
  HIF-2 alpha/EPAS1 antibody used for WB, ChIP, ICC-IF and IHC-P in experiments involving A498, 786-O, PANC-1, MIAPaCa-2, and ACHN cell lines and tumor microarray /TMAs comprising pretreatment nephrectomy specimens of patients with metastatic grade 3/4 CRCC treated with sorafenib or sorafenib plus interferon.
• Henze At, Garvalov Bk, Seidel S et al. Loss of PHD3 allows tumours to overcome hypoxic growth inhibition and sustain proliferation through EGFR. Nat Commun. 2014 Nov 25 [PMID: 25420773]
• Hou P, Kuo Cy, Cheng Ct et al. Intermediary Metabolite Precursor Dimethyl-2-Ketoglutarate Stabilizes Hypoxia-Inducible Factor-1Alpha by Inhibiting Prolyl-4-Hydroxylase PHD2. PLoS OnE. 2014 Nov 25 [PMID: 25420025] (WB, Human)
• Qi Y, Liu J, Saadat S et al. PTEn induces apoptosis and cavitation via HIF-2-dependent Bnip3 upregulation during epithelial lumen formation. Cell Death Differ. 2014 Nov 14 [PMID: 25394489] (WB, Mouse)
• Chen Sj, Hoffman Ne, Shanmughapriya S et al. A Splice Variant of the Human Ion Channel TRPM2 Modulates neuroblastoma Tumor Growth Through HIF-1/2Alpha. J. Biol. Chem. 2014 Nov 12 [PMID: 25391657] (WB, Human)
• Assar Me, Sanchez-Puelles Jm, Royo I et al. FM19G11 reverses endothelial dysfunction in rat and human arteries through stimulation of the PI3K/Akt/enOS pathway, independently of mTOR/HIF-1Alpha activation Br. J. Pharmacol. 2014 Nov 03 [PMID: 25363469] (WB, Rat)
  
  Details:
  HIF-1 alpha antibody used for WB on lysates of ex vivo FM19G11 treated aortic and mesenteric arteries from control and insulin-resistant rats (data not significant).
• Su Ej, Xin H, Yin P et al. Impaired fetoplacental angiogenesis in growth-restricted fetuses with abnormal umbilical artery Doppler velocimetry is mediated by aryl hydrocarbon receptor nuclear translocator (ARnT). J. Clin. Endocrinol. Metab. 2014 Oct 24 [PMID: 25343232] (WB, Human)
• Wu R, Chang Hc, Khechaduri A et al. Cardiac-specific ablation of ARnT leads to lipotoxicity and cardiomyopathy. J. Clin. Invest. 2014 Nov 03 [PMID: 25329697] (ChIP, Human)
• Hu H, Takano N, Xiang L et al. Hypoxia-inducible factors enhance glutamate signaling in cancer cells. Oncotarget 2014 Oct 15 [PMID: 25326682] (ChIP, WB, Human)
• Zhou Y, Yuge A, Rajah Am et al. LIMK1 Regulates Human Trophoblast Invasion/Differentiation and Is Down-Regulated in Preeclampsia. Am. J. Pathol. 2014 Oct 09 [PMID: 25307528] (IHC, Human)
• Nagaya Y, Aoyama M, Tamura T et al. Inflammatory cytokine tumor necrosis factor Alpha suppresses neuroprotective endogenous erythropoietin from astrocytes mediated by hypoxia-inducible factor-2Alpha. Eur. J. neurosci. 2014 Oct 04 [PMID: 25283246] (WB)
• Petruzzelli R, Christensen DR, Parry KL et al. HIF-2a Regulates NANOG Expression in Human Embryonic Stem Cells following Hypoxia and Reoxygenation through the Interaction with an Oct-Sox Cis Regulatory Element. PLoS ONE. 2014 Oct 02 [PMID: 25271810] (ICC/IF, ChIP, Human)
  
  Details:
  HIF-2 alpha/EPAS1 antibody used for ChIP and ICC-IF in experiments involving human embryonic stem cell /hESC.
• Bordji K, Grandval A, Cuhna-Alves L et al. Hypoxia-inducible factor-2a (HIF-2a), but not HIF-1a, is essential for hypoxic induction of class III b-tubulin expression in human glioblastoma cells. FEBS J. 2014 Sep 22 [PMID: 25244496] (WB, GS, Human)
  
  Details:
  HIF-2 alpha/EPAS1 antibody used for WB and EMSA/Gel Super Shift Assay in experiments involving human glioblastoma cell lines - U87MG (p53 wild-type, PTEN mutant), U251MG (p53 mutant, PTEN mutant) and GL15 (obtained from a frontal lobe tumor biopsy of a 59-year-old man with GBM).
• Muz B, de la Puente P, Azab F et al. Hypoxia Promotes Dissemination and Colonization in New Bone Marrow Niches in Waldenstrom's Macroglobulinemia. Mol. Cancer Res. 2014 Sep 17 [PMID: 25232031] (WB, Mouse)
• Lalevee S, Lapaire O, Buhler M. miR455 is linked to hypoxia signaling and is deregulated in preeclampsia. Cell Death Dis. 2014 Sep 05 [PMID: 25188518]
• Fujikuni N, Yamamoto H, Tanabe K et al. Hypoxia-mediated CD24 expression is correlated with gastric cancer aggressiveness by promoting cell migration and invasion. Cancer Sci 2014 Nov [PMID: 25174257] (WB)
• Asnaghi L, Lin MH, Lim KS et al. Hypoxia Promotes Uveal Melanoma Invasion through Enhanced Notch and MAPK Activation. PLoS ONE. 2014 Aug 28 [PMID: 25166211] (WB, Human)
• Yoshizawa Sayuri, Brown Andrew, Barchowsky Aaron, Sfeir Charles. Role of magnesium ions on osteogenic response in bone marrow stromal cells. Connect Tissue Res. 2014 Aug 01 [PMID: 25158202]
• Hubbi ME, Gilkes DM, Hu H et al. Cyclin-dependent kinases regulate lysosomal degradation of hypoxia-inducible factor 1alpha to promote cell-cycle progression. Proc Natl Acad Sci U S A. 2014 Jul 28 [PMID: 25071185] (WB, Human)
• Kurt B, Gerl K, Karger C et al. Chronic Hypoxia-Inducible Transcription Factor-2 Activation Stably Transforms Juxtaglomerular Renin Cells into Fibroblast-Like Cells In Vivo J Am Soc Nephrol. 2014 Jul 27 [PMID: 25071089] (IHC, Mouse)
• Li B, Qiu B, Lee DS et al. Fructose-1,6-bisphosphatase opposes renal carcinoma progression. Nature 2014 Jul 20 [PMID: 25043030] (WB, IP, Human)
• MEsange P, Poindessous V, Sabbah M et al. Intrinsic bevacizumab resistance is associated with prolonged activation of autocrine VEGF signaling and hypoxia tolerance in colorectal cancer cells and can be overcome by nintedanib, a small molecule angiokinase inhibitor Oncotarget. 2014 Jan 19 [PMID: 25015210] (IHC, WB, Mouse)
• Wong WJ, Richardson T, Seykora JT et al. Hypoxia-inducible factors regulate filaggrin expression and epidermal barrier function. J Invest Dermatol 2015 Feb [PMID: 24999590] (WB)
• Liu XD, Yao J, Tripathi DN et al. Autophagy mediates HIF2a degradation and suppresses renal tumorigenesis. Oncogene 2014 Jul 07 [PMID: 24998849] (WB, IP, Co-IP, Human)
  
  Details:
  HIF-2 alpha antibody used for WB, IP and CoIP in several experiments involving Caki-I cells, RPE cells, HEK293T stable cell line expressing HIF2a-GFP, 786-O parental cells/ and its over-expression derivatives, and Atg5+/+ or Atg5- / - MEFs stably expressing HIF2a-GFP.
• Slominski A, Kim TK, Brozyna AA et al. The role of melanogenesis in regulation of melanoma behavior: Melanogenesis leads to stimulation of HIF-1a expression and HIF-dependent attendant pathways. Arch. Biochem. Biophys. 2014 Jul 02 [PMID: 24997364] (WB, Human, Hamster)
  
  Details:
  HIF-2 alpha antibody used for WB on whole cell lysates of Bomirski hamster AbC1 and human SKMEL-188 stable lined (amelanotic /non-pigmented and melanotic /pigmented lines) with Caki cells as a positive control for HIF-2 alpha. Antibody used at 1:2000 dilution and the data is shown in Fig 1.
• Kono K, Tamashiro DA, Alarcon VB. Inhibition of RHO-ROCK signaling enhances ICM and suppresses TE characteristics through activation of hippo signaling in the mouse blastocyst. Dev. Biol. 2014 Jul 02 [PMID: 24997360] (WB, Human, Hamster)
  
  Details:
  HIF-2 alpha antibody used for WB on lysates of Bomirski hamster AbC1 and human SKMEL-188 cell lines (Fig. 1).
• Takano N, Sarfraz Y, Gilkes DM et al. Decreased Expression of Cystathionine b-Synthase Promotes Glioma Tumorigenesis. Mol. Cancer Res. 2014 Jul 03 [PMID: 24994751] (WB, Human)
  
  Details:
  HIF-2 alpha antibody used for WB on lysates of U87-MG subclones exposed to 1% or 20% O2 for 3 days and on HIF2 alpha depleted cells (Figure 5).
• Mangiavini L, Merceron C, Araldi E et al. Loss of VHL in mesenchymal progenitors of the limb bud alters multiple steps of endochondral bone development Dev. Biol. 2014 Jun 24 [PMID: 24972088] (IHC-P, Mouse)
  
  Details:
  HIF-2 alpha antibody used for IHC-P on forelimbs of E10.5, E12.5, E13.5 and E15.5 mice embryos - antigen retreival with sodium citrate buffer pH 6 at 95C for 10 min, primary dilution 1:100 ON/4C, detection via biotinylated secondary swine-anti-rabbit followed by labeled streptavidin biotin/TSA system (Perkin Elmer, USA), Negative controls - no primary antibody. (Images - Supplemental Fig. 1, Panel C - c/d)
• Rana S, Rajakumar A, Geahchan C et al. Ouabain inhibits placental sFlt1 production by repressing HSP27-dependent HIF-1alpha pathway. FASEB J. 2014 Jun 26 [PMID: 24970393] (WB, Human)
  
  Details:
  Used for WB on HIF-2 expression in both normal and pre-eclamptic placental villous explant cultures treated with 500 nM ouabain(Fig. 3F, G).
• Furcht CM, Buonato JM, Skuli N et al. Multivariate signaling regulation by SHP2 differentially controls proliferation and therapeutic response in glioma cells. J. Cell. Sci. 2014 Jun 20 [PMID: 24951116] (WB, Human)
  
  Details:
  Antibody used for detecting HIF2 alpha by WB in U87MG-M 281 control or SHP2 knockdown cells generated ectopic xenograft tumors in Nu/Nu mice (Fig. 6C, 6D)
• Salman Shaima, Holloway Alison C, Nurse Colin A. Chronic opioids regulate KATP channel subunit Kir6.2 and carbonic anhydrase I and II expression in rat adrenal chromaffin cells via HIF-2alpha and protein kinase A. Am J Physiol Cell Physiol. 2014 Aug 01 [PMID: 24898587] (Rat)
• Arreola A, Cowey CL, Coloff JL et al. HIF1alpha and HIF2alpha Exert Distinct Nutrient Preferences in Renal Cells. PLoS ONE. 2014 Jun 02 [PMID: 24879016] (ICC/IF, Mouse)
• Chaturvedi P, Gilkes DM, Takano N, Semenza GL. Hypoxia-inducible factor-dependent signaling between triple-negative breast cancer cells and mesenchymal stem cells promotes macrophage recruitment. Proc Natl Acad Sci U S A. 5/5/2014 [PMID: 24799675] (ChIP, Human)
• Ding Z, German P, Bai S et al. Genetic and Pharmacological Strategies to Refunctionalize the von Hippel Lindau R167Q Mutant Protein. Cancer Research 2014 Mar 13 [PMID: 24755468] (WB, Human)
• Gerovac BJ, Valencia M, Baumlin N et al. Submersion and Hypoxia Inhibit Ciliated Cell Differentiation in a Notch Dependent Manner.. Am. J. Respir. Cell Mol. Biol. [PMID: 24754775]
• Pang M, Liu X, Slagle-Webb B. Role of H-Ferritin in Radiosensitivity of Human Glioma Cells. HSOA Journal of Cancer Biology and Treatment Jan 21 2016 [PMID: 24707546] (WB, Human)
• Kihira Yoshitaka, Miyake Mariko, HiRata Manami et al. Deletion of hypoxia-inducible factor-1a in adipocytes enhances glucagon-like peptide-1 secretion and reduces adipose tissue inflammation. PloS One 2014 [PMID: 24705496] (WB, Mouse)
• Qin N, de Cubas AA, Garcia-Martin R et al. Opposing effects of HIF1a and HIF2a on chromaffin cell phenotypic features and tumor cell proliferation: Insights from MAX. Int. J. Cancer 3/27/2014 [PMID: 24676840] (IHC-P, WB, Human, Mouse)
• Heyboer M, Milovanova TN, Wojcik S et al. CD34+/CD45-dim stem cell mobilization by hyperbaric oxygen - Changes with oxygen dosage. Stem Cell Res 3/19/2014 [PMID: 24642336] (FLOW, Human)
• Zhang J, Ma WY. Nerve growth factor regulates the expression of vascular endothelial growth factor in human HaCaT keratinocytes via PI3K/mTOR pathway. Genet. Mol. Res. 1/24/2014 [PMID: 24615084] (WB, Human)
• Huang YH, Lin MH, Wang PC et al. Hypoxia inducible factor 2a/insulin-like growth factor receptor signal loop supports the proliferation and Oct-4 maintenance of mouse germline stem cells. Mol. Hum. Reprod. 3/23/2014 [PMID: 24598112] (ICC/IF, WB, Mouse)
• Lian G, Wei C, Wang D et al. Protein Profiling of H. pylori-Associated Gastric Cancer. Am. J. Pathol. 2014 Feb 28 [PMID: 24589339] (WB, Human)
• Fiorito Veronica, Neri Francesco, Pala Valentina et al. Hypoxia controls Flvcr1 gene expression in Caco2 cells through HIF2alpha and ETS1. Biochim Biophys Acta. 2014 Feb 24 [PMID: 24576667]
• Yoshizawa S, Brown A, Barchowsky A. Magnesium ion stimulation of bone marrow stromal cells enhances osteogenic activity, simulating the effect of magnesium alloy degradation. Acta Biomater 2014 Feb 24 [PMID: 24512978] (WB, Human)
• Duan LJ, Takeda K, Fong GH. Hematological, Hepatic, and Retinal Phenotypes in Mice Deficient for Prolyl Hydroxylase Domain (PHD) Proteins in the Liver. Am. J. Pathol. 2014 Feb 19 [PMID: 24508125] (WB, Mouse)
• Wojciak-Stothard B, Abdul-Salam Vb, Lao Kh et al. Aberrant chloride intracellular channel 4 expression contributes to endothelial dysfunction in pulmonary arterial hypertension Circulation et al. 2014 Apr 29 [PMID: 24503951] (WB, Mouse)
  
  Details:
  HIF2 alpha antibody used for WB on lysates from lungs of wildtype (WT) and CLIC4 KO mice subjected to normoxic and hypoxic conditions.
• Weisel FC, Kloepping C, Pichl A et al. Impact of S-Adenosylmethionine Decarboxylase 1 in Pulmonary Vascular Remodeling. Circulation 2014 Jan 28 [PMID: 24470481] (ChIP, Mouse, Human)
• Chen Z, Liu X, Mei Z, Xiao W. EAF2 Suppresses Hypoxia-Induced Factor 1 alpha Transcriptional Activity by Disrupting Its Interaction with Coactivator CBP/p300. Mol. Cell. Biol. 2014 Mar 1 [PMID: 24421387] (WB, Human)
• Kumar V, Everingham S, Hall C et al. Calpains promote neutrophil recruitment and bacterial clearance in an acute bacterial peritonitis model. Eur J Immunol 2013 Dec 5 [PMID: 24375267] (WB, Mouse)
• Place TL, Nauseef JT, Peterson MK et al. Prolyl-4-Hydroxylase 3 (PHD3) Expression Is Downregulated during Epithelial-to-Mesenchymal Transition. PLoS One 2013 Dec 18 [PMID: 24367580] (WB, Human)
• Gomes AP, Price NL, Ling AJ et al. Declining NAD+ Induces a Pseudohypoxic State Disrupting Nuclear-Mitochondrial Communication during Aging. Cell 2013 Dec 19 [PMID: 24360282] (WB, Mouse)
• Pasupuleti V, Du W, Gupta Y et al. Dysregulated D-Dopachrome Tautomerase, a Hypoxia Inducible Factor-Dependent Gene, Cooperates with Macrophage Migration Inhibitory Factor in Renal Tumorigenesis. J Biol Chem 2013 Dec 19 [PMID: 24356968] (ChIP, WB, Human)
• Zhu J, Jiang X, Chehab FF. FoxO4 Interacts with the Sterol Regulatory Factor SREBP2 and the Hypoxia Inducible Factor HIF2A at the CYP51 Promoter. J Lipid Res 2013 Dec 18 [PMID: 24353279] (WB, ChIP, Mouse)
• Razorenova OV, Castellini L, Colavitti R et al. The Apoptosis Repressor with a CARD Domain (ARC) Gene Is a Direct Hypoxia-Inducible Factor 1 Target Gene and Promotes Survival and Proliferation of VHL-Deficient Renal Cancer Cells. Mol Cell Biol 2014 Feb [PMID: 24344197] (WB, Human)
• Takano N, Peng YJ, Kumar GK et al. Hypoxia-inducible factors regulate human and rat cystathionine B-synthase gene expression. Biochem J 2013 Dec 12 [PMID: 24328859] (WB, ChIP, Human, Rat)
• Bocca C, Ievolella M, Autelli R et al. Expression of Cox-2 in human breast cancer cells as a critical determinant of epithelial-to-mesenchymal transition and invasiveness. Expert Opin Ther Targets 2014 Feb [PMID: 24325753] (IHC-P, Human)
• Chen Z, He X, Xia W et al. Prognostic Value and Clinicopathological Differences of HIFs in Colorectal Cancer: Evidence from Meta-Analysis. PLoS One 2013 Dec 6 [PMID: 24324596] (IHC, Human)
• Gilkes DM, Xiang L, Lee SJ et al. Hypoxia-inducible factors mediate coordinated RhoA-ROCK1 expression and signaling in breast cancer cells. Proc Natl Acad Sci U S A 2014 Jan 21 [PMID: 24324133] (ChIP, WB, Human)
• Chiatar SS, Eze OP, Schoenfeld AR. Expression of VHL Causes Three-Dimensional Morphological Changes in Renal Cells Indicative of Proximal Tubule Differentiation Cell Dev Biol. 2013 Aug 12 [PMID: 24308012] (WB, Human)
• Glover LE, Bowers BE, Saeedi B et al. Control of creatine metabolism by HIF is an endogenous mechanism of barrier regulation in colitis. Proc Natl Acad Sci U S A 2013 Dec 3 [PMID: 24248342] (ChIP, WB, Human)
• Xiang L, Gilkes DM, Chaturvedi P, Semenza GL et al. Ganetespib blocks HIF-1 activity and inhibits tumor growth, vascularization, stem cell maintenance, invasion, and metastasis in orthotopic mouse models of triple-negative breast cancer. J Mol Med (Berl). 2013 Nov 20 [PMID: 24248265]
• Proper SP, Saini Y, Greenwood KK et al. Loss of hypoxia-inducible factor 2 alpha (HIF2alpha) in the lung alveolar epithelium of mice leads to enhanced eosinophilic inflammation in cobalt-induced lung injury. Toxicol Sci. 2013 Nov 11 [PMID: 24218148] (IHC-P, Mouse)
• Xu Z, Zhao L, Zhu LY et al. MicroRNA-17, 20a Regulates the Proangiogenic Function of Tumor-Associated Macrophages via Targeting Hypoxia-Inducible Factor 2alpha. PLoS One. 2013 Oct 23 [PMID: 24194900] (IHC-P, IHC-Fr, ICC/IF, WB, Human)
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• Koizume, S et al. (2006) Activation of cancer cell migration and invasion by ectopic synthesis of coagulation factor VII. Cancer Res;66(19):9453-60. 2006 Oct 1. [PMID: 17018600] (WB, ChIP, Human)
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• Chavez JC, Baranova O, Lin J, Pichiule P. The transcriptional activator hypoxia inducible factor 2 (HIF-2/EPAS-1) regulates the oxygen-dependent expression of erythropoietin in cortical astrocytes. J Neurosci;26(37):9471-81. 2006 Sep 13. [PMID: 16971531] (WB, ELISA, IHC, ChIP, Mouse)
• Takeda, K et al. Placental but not heart defects are associated with elevated hypoxia-inducible factor alpha levels in mice lacking prolyl hydroxylase domain protein 2. Mol Cell Biol;26(22):8336-46. 2006 Nov. [PMID: 16966370] (WB, ELISA, Mouse)
• Carroll PA, Kenerson HL, Yeung RS, Lagunoff M. Latent Kaposi's sarcoma-associated herpesvirus infection of endothelial cells activates hypoxia-induced factors. J Virol;80(21):10802-12. 2006 Nov. [PMID: 16956952] (IHC-P, Human)
• Liu YL, Yu JM, Song XR et al. Regulation of the chemokine receptor CXCR4 and metastasis by hypoxia-inducible factor in non small cell lung cancer cell lines. Cancer Biol Ther 2006 Oct [PMID: 16929169] (FLOW, Human)
• Lin Q, Lee YJ, Yun Z. Differentiation arrest by hypoxia. J Biol Chem;281(41):30678-83. 2006 Oct 13. [PMID: 16926163]
• Shinojima, T et al. Renal cancer cells lacking hypoxia inducible factor (HIF)-1alpha expression maintain vascular endothelial growth factor expression through HIF-2alpha. Carcinogenesis;28(3):529-36. 2007 Mar. [PMID: 16920734] (IB, Human)
• Franovic, A et al. (2006) Multiple acquired renal carcinoma tumor capabilities abolished upon silencing of ADAM17. Cancer Res;66(16):8083-90. 2006 Aug 15. [PMID: 16912185] (WB, Human)
• Lutz MS, Burk RD. Primary cilium formation requires von hippel-lindau gene function in renal-derived cells. Cancer Res;66(14):6903-7. 2006 Jul 15. [PMID: 16849532] (WB, Human)
• Hergovich, A et al. Priming-dependent phosphorylation and regulation of the tumor suppressor pVHL by glycogen synthase kinase 3. Mol Cell Biol;26(15):5784-96. 2006 Aug. [PMID: 16847331]
• Bracken CP, Fedele AO, Linke S et al. Cell-specific regulation of hypoxia-inducible factor (HIF)-1alpha HIF-2alpha stabilization transactivation in a graded oxygen environment. J Biol Chem;281(32):22575-85. 2006 Aug 11. [PMID: 16760477] (WB, Human)
• Nakamura, K et al. (2006) Brain-derived neurotrophic factor activation of TrkB induces vascular endothelial growth factor expression via hypoxia-inducible factor-1alpha in neuroblastoma cells. Cancer Res;66(8):4249-55. 2006 Apr 15. [PMID: 16618748] (WB, IP, Human)
• Hu, C-J et al. Differential regulation of the transcriptional activities of hypoxia-inducible factor 1 alpha (HIF-1alpha) and HIF-2alpha in stem cells. Mol Cell Biol;26(9):3514-26. 2006 May. [PMID: 16611993]
• Ietta, F et al. Dynamic HIF1A regulation during human placental development. Biol Reprod;75(1):112-21. 2006 Jul. [PMID: 16611863] (WB, Human)
• Okuyama, H et al. (2006) Expression of vascular endothelial growth factor receptor 1 in bone marrow-derived mesenchymal cells is dependent on hypoxia-inducible factor 1. J Biol Chem;281(22):15554-63. 2006 Jun 2. [PMID: 16574650]
• Mizukami, Y et al. (2006) Hypoxic regulation of vascular endothelial growth factor through the induction of phosphatidylinositol 3-kinase/Rho/ROCK and c-Myc. J Biol Chem;281(20):13957-63. 2006 May 19. [PMID: 16543245] (WB, Human)
• Krishnamachary, B et al. Hypoxia-inducible factor-1-dependent repression of E-cadherin in von Hippel-Lindau tumor suppressor-null renal cell carcinoma mediated by TCF3, ZFHX1A, and ZFHX1B. Cancer Res;66(5):2725-31. 2006 Mar 1. [PMID: 16510593] (IB, Human)
• Hui, A S et al. Calcium signaling stimulates translation of HIF-alpha during hypoxia. FASEB J;20(3):466-75. 2006 Mar. [PMID: 16507764]
• Kurban, G et al. Characterization of a von Hippel Lindau pathway involved in extracellular matrix remodeling, cell invasion, and angiogenesis. Cancer Res;66(3):1313-9. 2006 Feb 1. [PMID: 16452184]
• Schultz, K, Fanburg, B L, Beasley, D. Hypoxia and hypoxia-inducible factor-1alpha promote growth factor-induced proliferation of human vascular smooth muscle cells. Am J Physiol Heart Circ Physiol; 290(6):H2528-34. 2006 Jun. [PMID: 16399861] (WB, Human)
• Calvani, M et al. Hypoxic induction of an HIF-1alpha-dependent bFGF autocrine loop drives angiogenesis in human endothelial cells. Blood;107(7):2705-12. 2006 Apr 1. [PMID: 16304044] (IB, Human)
• Lee S, Nakamura E, Yang H et al. Neuronal apoptosis linked to EglN3 prolyl hydroxylase and familial pheochromocytoma genes: developmental culling and cancer. Cancer Cell 2005 Aug [PMID: 16098468] (WB, Human)
• Zagzag D, Krishnamachary B, Yee H et al. Stromal cell-derived factor-1alpha and CXCR4 expression in hemangioblastoma and clear cell-renal cell carcinoma: von Hippel-Lindau loss-of-function induces expression of a ligand and its receptor. Cancer Res 2005 Jul 15 [PMID: 16024619] (IHC)
• Asikainen TM, Schneider BK, Waleh NS et al. Activation of hypoxia-inducible factors in hyperoxia through prolyl 4-hydroxylase blockade in cells and explants of primate lung. Proc Natl Acad Sci U S A;102(29):10212-7. 2005 Jul 19. [PMID: 16009933] (WB, Human)
• Maltepe E, Krampitz GW, Okazaki KM et al. Hypoxia-inducible factor-dependent histone deacetylase activity determines stem cell fate in the placenta. Development 2005 Aug [PMID: 15987772] (WB, Mouse)
• Smith, K et al. Silencing of epidermal growth factor receptor suppresses hypoxia-inducible factor-2-driven VHL-/- renal cancer. Cancer Res;65 (12):5221-30. 2005 Jun 15. [PMID: 15958567] (WB, Mouse)
• Temes, E et al. Activation of HIF-prolyl hydroxylases by R59949, an inhibitor of the diacylglycerol kinase. J Biol Chem;280(25):24238-44. 2005 Jun 24. [PMID: 15849364] (WB, Human)
• Miller, F et al. Inactivation of VHL by tumorigenic mutations that disrupt dynamic coupling of the pVHL. J Biol Chem.280(9):7985-96. 2005Mar 4. [PMID: 15611064] (WB, Human)
• Petrella BL, Lohi J, Brinckerhoff CE. Identification of membrane type-1 matrix metalloproteinase as a target of hypoxia-inducible factor-2 alpha in von Hippel-Lindau renal cell carcinoma. Oncogene 2005 Feb 3 [PMID: 15592504] (ELISA, Human)
• Rathmell, W et al. In vitro and in vivo models analyzing von Hippel-Lindau disease-specific mutations. Cancer Res;64(23):8595-603. 2004 Dec 1. [PMID: 15574766] (IB, IP, Mouse )
• Tang, N,et al. Loss of HIF-1alpha in endothelial cells disrupts a hypoxia-driven VEGF autocrine loop necessary for tumorigenesis. Cancer Cell;6 (5):485-95. 2004 Nov. [PMID: 15542432] (WB, Mouse)
• Freeburg, P et al. Divergent expression patterns for hypoxia-inducible factor-1beta and aryl hydrocarbon receptor nuclear transporter-2 in developing kidney. J Am Soc Nephrol;15(10):2569-78. 2004 Oct. [PMID: 15466261] (WB, Mouse)
• Biju MP, Neumann AK, Bensinger SJ, Johnson RS, Turka LA, Haase VH. Vhlh gene deletion induces Hif-1-mediated cell death in thymocytes. Mol Cell Biol;24(20):9038-47. 2004 Oct. [PMID: 15456877] (WB, Mouse)
• Rajakumar A, Brandon HM, Daftary A et al. Evidence for the functional activity of hypoxia-inducible transcription factors overexpressed in preeclamptic placentae. Placenta 2004 Nov [PMID: 15451190] (WB, Human)
• Takeda, N et al. Endothelial PAS domain protein 1 gene promotes angiogenesis through the transactivation of both vascular endothelial growth factor and its receptor, Flt-1. Circ Res;95(2):146-53. 2004 Jul 23. [PMID: 15192019] (WB)
• Grabmaier, K et al. Strict regulation of CAIX(G250/MN) by HIF-1alpha in clear cell renal cell carcinoma. Oncogene;23(33):5624-31. 2004 Jul 22. [PMID: 15184875] (GS, Human)
• Mekhail, K et al. HIF activation by pH-dependent nucleolar sequestration of VHL. Nat Cell Biol;6(7):642-7. 2004 Jul. [PMID: 15181450]
• Pan, Y et al. p53 cannot be induced by hypoxia alone but responds to the hypoxic microenvironment. Oncogene;23(29):4975-83. 2004 Jun 24. [PMID: 15107830] (WB, Mouse)
• Sato, M et al. The PAI-1 gene as a direct target of endothelial PAS domain protein-1 in adenocarcinoma A549 cells. Am J Respir Cell Mol Biol;31 (2):209-15. 2004 Aug. [PMID: 15039136]
• Welsh, S et al. Antitumor activity and pharmacodynamic properties of PX-478, an inhibitor of hypoxia-inducible factor-1alpha. Mol Cancer Ther;3(3):233-44. 2004 Mar. [PMID: 15026543] (IHC-P, Mouse)
• Mizukami, Y et al. Hypoxia-inducible factor-1-independent regulation of vascular endothelial growth factor by hypoxia in colon cancer. Cancer Res;64 (5):1765-72. 2004 Mar 1. [PMID: 14996738] (WB, Human)
• Zimmer, M et al. Inhibition of hypoxia-inducible factor is sufficient for growth suppression of VHL-/- tumors. Mol Cancer Res;2(2):89-95. 2004 Feb. [PMID: 14985465] (WB, Human)
• Uchida, T et al. Prolonged hypoxia differentially regulates hypoxia-inducible factor (HIF)-1alpha and HIF-2alpha expression in lung epithelial cells: implication of natural antisense HIF-1alpha. J Biol Chem;279(15):14871-8. 2004 Apr 9. [PMID: 14744852] (IB, Human)
• Martin-Puig, S et al. Role of iron (II)-2-oxoglutarate-dependent dioxygenases in the generation of hypoxia-induced phosphatidic acid through HIF-1/2 and von Hippel-Lindau-independent mechanisms. J Biol Chem;279(10):9504-11. 2004 Mar 5. [PMID: 14681229] (WB, Human)
• Hu CJ, Wang LY, Chodosh LA et al. Differential roles of hypoxia-inducible factor 1alpha (HIF-1alpha) and HIF-2alpha in hypoxic gene regulation. Mol Cell Biol 2003 Dec [PMID: 14645546] (WB)
• Yuan HT, Li XZ, Pitera JE et al. Peritubular capillary loss after mouse acute nephrotoxicity correlates with down-regulation of vascular endothelial growth factor-A and hypoxia-inducible factor-1 alpha. Am J Pathol 2003 Dec [PMID: 14633603] (IHC-P, WB, Mouse)
• Cuevas Y, Hernandez-Alcoceba R, Aragones J, Naranjo-Suarez S, Castellanos MC, Esteban MA, Martin-Puig S, Landazuri MO, del Peso L. Specific oncolytic effect of a new hypoxia-inducible factor-dependent replicative adenovirus on von Hippel-Lindau-defective renal cell carcinomas. Cancer Res;63(20):6877-84. 2003 Oct 15. [PMID: 14583486] (WB, Human)
• Corn PG, McDonald ER 3rd, Herman JG, El-Deiry WS. Tat-binding protein-1, a component of the 26S proteasome, contributes to the E3 ubiquitin ligase function of the von Hippel-Lindau protein. Nat Genet;35(3):229-37. 2003 Nov. [PMID: 14556007]
• Peso, L et al. The von Hippel Lindau/hypoxia-inducible factor (HIF) pathway regulates the transcription of the HIF-proline hydroxylase genes in response to low oxygen. J Biol Chem;278(49):48690-5. 2003 Dec 5. [PMID: 14506252] (WB, Human)
• Staller, P et al. Chemokine receptor CXCR4 downregulated by von Hippel-Lindau tumour suppressor pVHL. Nature;425(6955):307-11. 2003 Sep 18. [PMID: 13679920] (WB, Human)
• Mohan, S et al. von Hippel-Lindau protein complex is regulated by cell density. Oncogene;22(34):5270-80. 2003 Aug 14. [PMID: 12917628] (WB, Human)
• Palayoor, S et al. Ibuprofen-mediated reduction of hypoxia-inducible factors HIF-1alpha and HIF-2alpha in prostate cancer cells. Clin Cancer Res;9 (8):3150-7. 2003 Aug 1. [PMID: 12912967] (WB, Human)
• Mabjeesh, N et al. Androgens stimulate hypoxia-inducible factor 1 activation via autocrine loop of tyrosine kinase receptor/phosphatidylinositol 3'-kinase/protein kinase B in prostate cancer cells. Clin Cancer Res;9(7):2416-25. 2003 Jul. [PMID: 12855613] (WB, Human)
• Park, S et al. Hypoxia-induced gene expression occurs solely through the action of hypoxia-inducible factor 1alpha (HIF-1alpha): role of cytoplasmic trapping of HIF-2alpha. Mol Cell Biol;23(14):4959-71. 2003 Jul. [PMID: 12832481] (WB, ICC/IF, Mouse)
• Naranjo-Suarez, S et al. Down-regulation of hypoxia-inducible factor-2 in PC12 cells by nerve growth factor stimulation. J Biol Chem;278(34):31895-901. 2003 Aug 22. [PMID: 12805361] (WB, Rat)
• Liu MY, Poellinger L, Walker CL. Up-regulation of hypoxia-inducible factor 2alpha in renal cell carcinoma associated with loss of Tsc-2 tumor suppressor gene. Cancer Res 2003 May 15 [PMID: 12750296] (Rat)
• Baba M, Hirai S, Yamada-Okabe H et al. Loss of von Hippel-Lindau protein causes cell density dependent deregulation of CyclinD1 expression through hypoxia-inducible factor. Oncogene 2003 May 8 [PMID: 12743597] (WB, Human)
• Mabjeesh, N et al. 2ME2 inhibits tumor growth and angiogenesis by disrupting microtubules and dysregulating HIF. Cancer Cell;3(4):363-75. 2003 Apr. [PMID: 12726862] (WB, Human)
• Khatua, S et al. Overexpression of the EGFR/FKBP12/HIF-2alpha pathway identified in childhood astrocytomas by angiogenesis gene profiling. Cancer Res;63(8):1865-70. 2003 Apr 15. [PMID: 12702575] (IHC-P, Human)
• Graven,K et al. HIF-2alpha regulates glyceraldehyde-3-phosphate dehydrogenase expression in endothelial cells. Biochim Biophys Acta;1626(1-3):10-8. 2003 Apr 15. [PMID: 12697324]
• Toro, R et al. Induction of T-type calcium channel gene expression by chronic hypoxia. J Biol Chem;278(25):22316-24. 2003 Jun 20. [PMID: 12679337] (WB, Rat)
• Yuan, Y et al. Cobalt inhibits the interaction between hypoxia-inducible factor-alpha and von Hippel-Lindau protein by direct binding to hypoxia-inducible factor-alpha. J Biol Chem;278(18):15911-6. 2003 May 2. [PMID: 12606543] (WB, Rat)
• Mack, F et al. Loss of pVHL is sufficient to cause HIF dysregulation in primary cells but does not promote tumor growth. Cancer Cell;3(1):75-88. 2003 Jan. [PMID: 12559177] (WB, Mouse)
• Chan DA, Sutphin PD, Denko NC, Giaccia AJ. Role of prolyl hydroxylation in oncogenically stabilized hypoxia-inducible factor-1alpha. J Biol Chem;277(42):40112-7. 2002 Oct 18. [PMID: 12186875] (WB, Human)
• Bernaudin M, Tang Y, Reilly M, Petit E, Sharp FR. Brain genomic response following hypoxia and re-oxygenation in the neonatal rat. Identification of genes that might contribute to hypoxia-induced ischemic tolerance. J Biol Chem. 277(42):39728-38. 2002 Oct 18. [PMID: 12145288] (WB, Rat)
• Maranchie, J et al. The contribution of VHL substrate binding and HIF1-alpha to the phenotype of VHL loss in renal cell carcinoma. Cancer Cell;1(3):247-55. 2002 Apr. [PMID: 12086861] (WB, Human)
• Kondo, K et al. Inhibition of HIF is necessary for tumor suppression by the von Hippel-Lindau protein. Cancer Cell;1(3):237-46. 2002 Apr. [PMID: 12086860] (WB, Human)
• Xia, G et al. Positive expression of HIF-2alpha/EPAS1 in invasive bladder cancer. Urology;59(5):774-8. 2002 May. [PMID: 11992927] (ChIP, WB, Human)
• Liang, Y et al. Activation of vascular endothelial growth factor A transcription in tumorigenic glioblastoma cell lines by an enhancer with cell type-specific DNase I accessibility. J Biol Chem;277(22):20087-94. 2002 May 31. [PMID: 11912213]
• Alvarez-Tejado M, Alfranca A, Aragones J et al. Lack of evidence for the involvement of the phosphoinositide 3-kinase/Akt pathway in the activation of hypoxia-inducible factors by low oxygen tension. J Biol Chem;277(16):13508-17. 2002 Apr 19. [PMID: 11815624] (WB, Human)
• Burke B, Tang N, Corke KP, Tazzyman D, Ameri K, Wells M, Lewis CE. Expression of HIF-1alpha by human macrophages: implications for the use of macrophages in hypoxia-regulated cancer gene therapy. J Pathol;196(2):204-12. 2002 Feb. [PMID: 11793372]
• Michenko, A et al. Hypoxia-inducible factor-1-mediated expression of the 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase-3 (PFKFB3) gene. Its possible role in the Warburg effect. J Biol Chem. 277(8):6183-7. 2002 Feb 22. [PMID: 11744734]
• Makino, Y et al. Inhibitory PAS domain protein is a negative regulator of hypoxia-inducible gene expression. Nature;414(6863):550-4. 2001 Nov 29. [PMID: 11734856]
• Ad N, Snir E, Vidne BA, Golomb E. Histologic atrial myolysis is associated with atrial fibrillation after cardiac operation. Ann Thorac Surg;72(3):688-93. 2001 Sep. [PMID: 11565642] (IHC-P, Human)
• Luo JC, Shibuya M. A variant of nuclear localization signal of bipartite-type is required for the nuclear translocation of hypoxia inducible factors (1alpha, 2alpha and 3alpha). Oncogene;20(12):1435-44. 2001 Mar 22. [PMID: 11313887]
• Alvarez-Tejado M, Naranjo-Suarez S, Jimenez C et al. Hypoxia induces the activation of the phosphatidylinositol 3-kinase/Akt cell survival pathway in PC12 cells: protective role in apoptosis. J Biol Chem;276(25):22368-74. 2001 Jun 22. [PMID: 11294857] (WB, Mouse)
• Haase, VH et al. Vascular tumors in livers with targeted inactivation of the von Hippel-Lindau tumor suppressor. Proc Natl Acad Sci U S A;98 4):1583-8. 2001 Feb 13. [PMID: 11171994]
• de Paulsen N, Brychzy A, Fournier MC, Klausner RD, Gnarra JR, Pause A, Lee S. Role of transforming growth factor-alpha in von Hippel--Lindau (VHL)(-/-) clear cell renal carcinoma cell proliferation: a possible mechanism coupling VHL tumor suppressor inactivation and tumorigenesis. Proc Natl Acad Sci U S A;98(4):1387-92. 2001 Feb 13. [PMID: 11171960] (WB, FLOW, Human)
• Bonicalzi ME, Groulx I, de Paulsen N, Lee S. Role of exon 2-encoded beta -domain of the von Hippel-Lindau tumor suppressor protein. J Biol Chem;276(2):1407-16. 2001 Jan 12. [PMID: 11024059] (WB, IP, Human)
• Rajakumar, A, Conrad, K. Expression, ontogeny, and regulation of hypoxia-inducible transcription factors in the human placenta. Biol Reprod;63 (2):559-69. 2000 Aug. [PMID: 10906065] (WB, IHC-P, Human)
• Ohh, M et al. Ubiquitination of hypoxia-inducible factor requires direct binding to the beta-domain of the von Hippel-Lindau protein. Nat Cell Biol;2(7):423-7. 2000 Jul. [PMID: 10878807]
• Conrad PW, Freeman TL, Beitner-Johnson D et al. EPAS1 trans-activation during hypoxia requires p42/p44 MAPK. J Biol Chem;274(47):33709-13. 1999 Nov 19. [PMID: 10559262] (WB, IP, Rat)
• Baba M, Hirai S, Yamada-Okabe H et al. Loss of von Hippel-Lindau protein causes cell density dependent deregulation of CyclinD1 expression through hypoxia-inducible factor. Oncogene;22(18):2728-38. 2003 May 8. [PMID: 1274359] (WB, Human)

Reviews for HIF-2 alpha/EPAS1 Antibody (NB100-122) (31)

Average Rating: 4.4

(Based on 31 reviews)

We have 31 reviews tested in 4 species: Human, Mouse, Rat, Other.

reviewed by: Anonymous WB Human 07/05/2019

Summary

• Application: Western Blot
• Sample Tested: Human cancer cell lines
• Species: Human
• Lot: CF

reviewed by: Anonymous WB Human 04/16/2019

Summary

• Application: Western Blot
• Sample Tested: HBE-16
• Species: Human
• Lot: CD-9

reviewed by: Anonymous ChIP Human 12/02/2018

Summary

• Application: Chromatin Immunoprecipitation
• Sample Tested: MCF7 cell line
• Species: Human
• Lot: CD-13

reviewed by: Anonymous WB Human 09/18/2018

Summary

• Application: Western Blot
• Sample Tested: Human Cancer Cells
• Species: Human
• Lot: N/A

reviewed by: Anonymous WB Human 05/11/2018

Summary

• Application: Western Blot
• Sample Tested: 293T HEK cell lysate
• Species: Human
• Lot: CD-2

Comments

• Comments: Antibody dilution- 1:500 in 5% BSA- overnight 4 degerees

reviewed by: Shanhai Xie WB Mouse 01/12/2017

Summary

• Application: Western Blot
• Sample Tested: transfected U2OS cell lysate
• Species: Mouse
• Lot: BV-2

reviewed by: Anonymous WB Human 01/12/2017

Summary

• Application: Western Blot
• Sample Tested: cultured cells
• Species: Human
• Lot: BV-2

reviewed by: Anonymous WB Mouse 10/18/2016

Summary

• Application: Western Blot
• Sample Tested: Mouse CD4+ T Cells
• Species: Mouse
• Lot: BP-1

Comments

• Comments: Dilution (1:1000)

reviewed by: Helder Andre WB Human 07/11/2016

Summary

• Application: Western Blot
• Sample Tested: Retinal and Choroidal primary endothelia
• Species: Human

Blocking

• Blocking Details: 5% milk/TBS, 1h, RT

Primary Anitbody

• Dilution Ratio: 1:250 in 1% milk/TBS, ON, 4C

Secondary Antibody

• Secondary Description: swine anti-rabbit IgG-HRP
• Secondary Concentration: 1:5000 in 1% milk/TBS, 1h, RT

Details

• Detection Notes: ECL+

reviewed by: Anonymous WB Human 03/31/2016

Summary

• Application: Western Blot
• Sample Tested: HCT116 cell line lysate
• Species: Human
• Lot: BP-2

Blocking

• Blocking Details: 5% skim milk in TBS

Primary Anitbody

• Dilution Ratio: 1000 times, overnight incubation at 4 degrees C in 0.1% Tween-20 TBS 5% milk

Secondary Antibody

• Secondary Description: GE Healthcare Anti-rabbit IgG HRP
• Secondary Manufacturer Cat#: NA934V
• Secondary Concentration: 5,000 dilution

Details

• Detection Notes: Detection using autoradiography film, exposure 1 minutes, 24 hours hypoxia as positive controls.

reviewed by: Anonymous WB Human 11/10/2015

Summary

• Application: Western Blot
• Sample Tested: cytoplasmic and nuclear protein fractions of U266 cells
• Species: Human

Blocking

• Blocking Details: 5% milk in PBST, 30 minutes at Room Temperature

Primary Anitbody

• Dilution Ratio: 1 ug/ml, Overnight, 4 C, PBST with 5%milk

Secondary Antibody

• Secondary Description: Goat anti-rabbit igG HRP
• Secondary Manufacturer Cat#: SC-2005
• Secondary Concentration: 1:2000

Details

• Detection Notes: Chemiluminescence, G:Box Chemi XLT4, 12 sec, no positive and negative controls

Comments

• Comments: This antibody showed clean bands by wet transfer method.

reviewed by: Anonymous WB Human 08/12/2015

Summary

• Application: Western Blot
• Sample Tested: Primary Epithelial Cells
• Species: Human

Blocking

• Blocking Details: 5% Milk in 0.1% TBST for 1 hour at room temperature

Primary Anitbody

• Dilution Ratio: 1:500, Overnight at 4 celcius in 5% milk in 0.1% TBST

Secondary Antibody

• Secondary Description: Goat anti-rabbit HRP
• Secondary Manufacturer Cat#: KPL 0741516
• Secondary Concentration: 1:5000

Details

• Detection Notes: Chemiluminescence, BioRad Universal Hood II, 10 minute exposure

reviewed by: Anonymous WB Human 06/01/2015

Summary

• Application: Western Blot
• Sample Tested: MDA-MB-231 Cell Lysate
• Species: Human
• Lot: BQ1

Blocking

• Blocking Details: 5% milk

Primary Anitbody

• Dilution Ratio: 1:1000

Secondary Antibody

• Secondary Description: donkey anti-rabbit IgG, HRP
• Secondary Manufacturer Cat#: Amersham NA934V
• Secondary Concentration: 1:3000

Details

• Detection Notes: BIO RAD clarity western ECL (Antibody specificity; not react to HIF1a-overexpression as shown in the picture)

reviewed by: Anonymous WB Human 03/27/2015

Summary

• Application: Western Blot
• Sample Tested: 786O cell lysates
• Species: Human

Blocking

• Blocking Details: 5% milk in TBST

Primary Anitbody

• Dilution Ratio: 1:1000

Secondary Antibody

• Secondary Description: Goat anti-rabbit
• Secondary Concentration: 1:5000

Details

• Detection Notes: using the regular pico from pierece, detection by Bio-rad imager, 2 min exposure time, 786O vector cells is the positive ctrl.

reviewed by: Anonymous ChIP Human 12/12/2014

Summary

• Application: Chromatin Immunoprecipitation
• Sample Tested: See PMID:24248342
• Species: Human

Details

• Detection Notes: See PMID:24248342

Comments

• Comments: Published in PMID:24248342

reviewed by: Anonymous WB Human 12/12/2014

Summary

• Application: Western Blot
• Sample Tested: See PMID:24308012
• Species: Human

Blocking

• Blocking Details: See PMID:24308012

Details

• Detection Notes: Antibody works well but long exposure times are needed.

Comments

• Comments: Published in PMID:24308012

reviewed by: Xiuquan Luo WB Human 09/12/2014

Summary

• Application: Western Blot
• Sample Tested: HEK293 cell lysate
• Species: Human

Blocking

• Blocking Details: 1xPBST with 5% non-fat milk, RT 1 hour

Primary Anitbody

• Dilution Ratio: 1xPBST with 5% non-fat milk, overnight 4C, 1:1000 dilution

Secondary Antibody

• Secondary Description: Goat anti-rabbit IgG-HRP
• Secondary Manufacturer Cat#: sc-2004
• Secondary Concentration: 1:2000 dilution

Details

• Detection Notes: ECL dura, exposured for 2-5 min

reviewed by: Pramod Mallikarjuna WB Human 08/30/2014

Summary

• Application: Western Blot
• Sample Tested: Clear cell renal cell carcinoma samples
• Species: Human

Blocking

• Blocking Details: Odyssey Blocking Buffer

Primary Anitbody

• Dilution Ratio: 1:1000 in TBST, overnight, +4 degree celsius

Secondary Antibody

• Secondary Description: IRDye® 800CW Goat anti-Rat IgG
• Secondary Manufacturer Cat#: 926-32219
• Secondary Concentration: 1:10000

Details

• Detection Notes: Odyssey® CLx Infrared Imaging System

reviewed by: Anonymous WB Human 06/30/2014

Summary

• Application: Western Blot
• Sample Tested: Whole cell lysate
• Species: Human

reviewed by: Anonymous WB Human 04/30/2014

Summary

• Application: Western Blot
• Sample Tested: whole cell lysate U87MG (human glioblastoma cells)
• Species: Human

Blocking

• Blocking Details: 3% milk/TBST, 1h, room temperature

Primary Anitbody

• Dilution Ratio: 1:1000 in 3% milk/TBST, 2h, room temperature

Secondary Antibody

• Secondary Description: goat anti-rabbit IgG, HRP
• Secondary Manufacturer Cat#: Dako
• Secondary Concentration: 1:2000

Details

• Detection Notes: ECL, X-ray film, 1h exposure

reviewed by: Saori Yoshii IHC-P Mouse 04/11/2014

Summary

• Application: Immunohistochemistry-Paraffin
• Sample Tested: Mouse Intestine
• Species: Mouse
• Lot: BP-1
• Comments: HIF-2a can be detected in the nucleus. Background in the cytosol is also detected (HIF-2a real band only appeared in the nuclear fraction, checked by WB).

Blocking

• Blocking Details: Invitrogen HistoMouse kit

Primary Anitbody

• Dilution Ratio: x300 in 1% BSA

Secondary Antibody

• Secondary Description: Invitrogen HistoMouse kit

Details

• Detection Notes: DAB
• Fixation Details: +antigen retrieval

Comments

• Comments: HIF-2a can be detected in the nucleus. Background in the cytosol is also detected (HIF-2a real band only appeared in the nuclear fraction, checked by WB).

reviewed by: Anonymous WB Human 04/03/2014

Summary

• Application: Western Blot
• Species: Human
• Lot: BP-1

reviewed by: Prabir Chakraborty WB Human 01/24/2014

Summary

• Application: Western Blot
• Sample Tested: UNTREATED OVARIAN SURFACE EPITHELIUM CELLS (OSE) AND A2780 CELLS LYSATES 25ug
• Species: Human
• Lot: BB1

Blocking

• Blocking Details: 4% BSA IN TBST, 1H ROOM TEMPERATURE

Primary Anitbody

• Dilution Ratio: 1:750 IN 4% BSA IN TBST FOR O/N AT 4*C

Secondary Antibody

• Secondary Description: Goat anti-rabbit IgG-HRP
• Secondary Manufacturer Cat#: sc-2301
• Secondary Concentration: 1:10,000

Details

• Detection Notes: ECL

reviewed by: Anonymous WB Human 11/20/2013

Summary

• Application: Western Blot
• Sample Tested: Human Glioma cells, whole cell lysates
• Species: Human
• Lot: BN-1

Blocking

• Blocking Details: Milk 5% in Tween TBS, 30 min, 25°C

Primary Anitbody

• Dilution Ratio: 1/500, 16 hours at 4°C in Milk 1% in TBS Tween 0.1%

Secondary Antibody

• Secondary Description: Goat Anti-rabbit IgG (H+L)
• Secondary Manufacturer Cat#: Bio-Rad
• Secondary Concentration: 1/10000

Details

• Detection Notes: ECL

reviewed by: Anonymous IF Mouse 04/15/2013

Summary

• Application: Immunofluorescence
• Species: Mouse
• Pub Med ID: 22753893
• File: View PDF

reviewed by: Anonymous ChIP Rat 04/02/2013

Summary

• Application: Chromatin Immunoprecipitation
• Species: Rat
• Special Applications: Rat duodenal enterocyte

reviewed by: Anonymous WB 09/19/2012

Summary

• Application: Western Blot
• Lot: BK-2
• Pub Med ID: 21288809
• File: View PDF

reviewed by: Anonymous WB Mouse 12/08/2011

Summary

• Application: Western Blot
• Sample Tested: Mouse
• Species: Mouse

Blocking

• Blocking Details: 1Blocking Buffer: 0% Milk

Primary Anitbody

• Dilution Ratio: Dilution Ratio: 1:1000, Incubation Time: overnight, Incubation Temp: 4 degrees Celsius

Secondary Antibody

• Secondary Description: Secondary Ab Dilution Ratio: 1:5000

Details

• Detection Notes: Detection Method: ECL

reviewed by: Olga Razorenova WB Human 06/15/2011

Summary

• Application: Western Blot
• Sample Tested: ACHN, SN12C, Caki1 whole cell lysates, Sample Amount: 30ug
• Species: Human
• Lot: BC3
• Comments: Antibody at 1:500 dilution detects HIF2 alpha at the appropriate molecular weight (around 94kDa) in cells exposed to hypoxia (0.5% O2, 16h). No need to do nuclear extracts, it works for total cell protein extracts, also the amount of protein for lane should be 30-80ug. Usually 30 ug is enough.
  
  
  
  
  
  Good luck!!!
• Pub Med ID: 21233420

Blocking

• Blocking Details: Blocking Buffer: 5% Milk in PBST, Blocking Time: 1 hour, Blocking Temp: Room temperature

Primary Anitbody

• Dilution Ratio: Dilution Ratio: 1:500, Incubation Dilution Buffer: 5% Milk in PBST, Incubation Time: overnight, Incubation Temp: 4 C

Secondary Antibody

• Secondary Description: Secondary Ab: anti-Rabbit-HRP from Pierce, Secondary Ab Dilution Ratio: 1:5000
• Secondary Manufacturer Cat#: 31460

Details

• Detection Notes: Method: ECL, Exposure Time: 3 min, Pos.Ctrl: Hypoxia treatment, Neg.Ctrl: Normoxia (21% O2) treatment, Specific Bands: 94 kDa

Comments

• Comments: Antibody at 1:500 dilution detects HIF2 alpha at the appropriate molecular weight (around 94kDa) in cells exposed to hypoxia (0.5% O2, 16h). No need to do nuclear extracts, it works for total cell protein extracts, also the amount of protein for lane should be 30-80ug. Usually 30 ug is enough. Good luck!!!

reviewed by: Anonymous WB Other 03/11/2010

Summary

• Application: Western Blot
• Sample Tested: Colon Cancer Cell llines, Sample Amount: 50ug
• Species: Other
• Lot: AT1
• Comments: The image was of RKO cells. Using HCT116 cells we saw multiple bands, but the band at 118 was deinitely stronger. We image out westerns using a Tyhpoon and ECL Plus. We find we can use more dilute antibodies than when we were using regular ECL and film.

Blocking

• Blocking Details: Blocking Buffer: 5% Blotto, Blocking Time: 1 hour, Blocking Temp: Room temperature

Primary Anitbody

• Dilution Ratio: Dilution Ratio: 1/2000, Incubation Dilution Buffer: 5% BSA in TNS with Tween, Incubation Time: overnight, Incubation Temp: 4 C

Secondary Antibody

• Secondary Description: Secondary Ab: anti-Rabbit, Secondary Ab Dilution Ratio: 1/10000

Details

• Detection Notes: Detection Method: ECL Plus, Specific Bands: 118 kDa

Comments

• Comments: The image was of RKO cells. Using HCT116 cells we saw multiple bands, but the band at 118 was deinitely stronger. We image out westerns using a Tyhpoon and ECL Plus. We find we can use more dilute antibodies than when we were using regular ECL and film.

reviewed by: Anonymous WB Other 02/09/2009

Summary

• Application: Western Blot
• Sample Tested: head and neck squamous cell carcinoma cells, Sample Amount: 35 ug
• Species: Other

Blocking

• Blocking Details: Blocking Buffer: 5% nonfat dry milk in TBS-T, Blocking Time: 1 hour, Blocking Temp: 25 degrees Celsius

Primary Anitbody

• Dilution Ratio: Dilution Ratio: 1/500, Incubation Dilution Buffer: 5% nonfat dry milk in TBS-T, Incubation Time: overnight, Incubation Temp: 4 C

Secondary Antibody

• Secondary Description: Secondary Ab: rabbit polyclonal HRP, Secondary Ab Dilution Ratio: 1/10000

Details

• Detection Notes: Detection Method: ECL, Exposure Time: varies, Pos.Ctrl: hypoxia-treated sample, Neg.Ctrl: normoxia, Specific Bands: ~120 kDa

FAQs for HIF-2 alpha/EPAS1 Antibody (NB100-122). (Showing 1 - 10 of 10 FAQs).

1. What is the synthetic peptide of NB100-122, corresponding to which amino acids? Can this antibody cross-react with chicken or not?
   • Unfortunately we cannot provide any further information on this epitope as it is considered proprietary. The immunogen is a peptide derived from the C-terminus of mouse/human HIF-2 alpha protein. I have carried out a blast for the % homology between human HIF-2 alpha and Chicken HIF-2 alpha. The % homology is 75% and our lab only recommends use when the homology is 85% or above.
2. What do you recommend for blocking for a Western blot with this antibody?
   • We recommend to block membranes for 1.5 hours with 1X western wash buffer containing 5% non-fat dry milk (NFDM). For more information, a product specific WB protocol can be found on the datasheet if you click on the "Protocols & FAQs" tab.
3. We are using HIF2 Antibody from Novus to detect HIF2 in hypoxic HUVEC cells. What washing buffer do you use for Western Blot with HIF2? Are you using PBS or TBS, 0.3 or 0.1% Tween to solute the 5% milk (NFDM)? Do you solute the antibody 1:500 or 1:1000?
   • Please see our specific protocol for Hif2 here. We recommend 0.5 to 1.0% Tween as a washing buffer and we recommend a dilution of 1:200 to 1:1000. This is depending on your sample and you will have to optimize this accordingly.
4. I am working on a WB using the antibody against mouse HIF1a and HIF2a (catalog numbers NB100-105 and NB100-122). I tried several anti-mouse and anti-rabbit HRP secondary antibodies (from another company) and found they gave different patterns. Please provide the secondary antibody info which was used in the WB showed on the datasheet.
   • We use catalog numbers NB7570 and NB7183 as our secondary antibodies for testing these antibodies in WB.
5. Is this antibody useful for EMSA-supershift?
   • Unfortunately we have not tested if this antibody can be used in EMSA-supershift. If you would be interested in testing this novel application, please take a look at our Innovator's Reward program.
6. Will the hif protein be degraded when preparing the nuclear extract?
   • Working with Hif-2 can be tricky at times and the possibility of degradation is a very real one. Please see this link for a specific protocol for using NB100-122. The nuclear extract protocol can be found using this reference: Wang and Semenza. Purification and Characterization of Hypoxia-Inducible Factor 1. Journal of Biological Chemistry. 270(3): 1230-1237, 1995. I would also recommend that you use a positive control; a nuclear extract of hypoxia induced cell lines (293, Hep3B, COS7, Hepa) are all excellent choices. You can purchase the hypoxic COS7 lysate from us to make it more convenient - the catalog number is NB800-PC26. Please also see our Hypoxia support page for suggestions.
7. We've purchased NB100-122 for Western blot and IHC. I'd like to know what would be good positive control for this antibody in case of IHC.
   • For HIF2 alpha positive control tissues, we recommend the following: Kidney, Ling, Brain, Heart. For western blot, we recommend using a positive and negative control such as normoxic/hypoxic or treated/untreated with CoCl2. We sell a CoCl2 treated/untreated Cos7 lysate: View Cos7 lysate datasheet.
8. What is the sequence for the peptide used to purify this antibody?
   • The exact sequence of the immunogen is proprietary, but I can tell you the range is within 625-695.
9. Could you tell us if these antibodies need to use cell lysate treated with special extraction for WB? Do you have the recommended preparation method of cell lysate?
   • Usually HIF1 alpha and HIF2 alpha are hard to detect endogenously and require some inducement. The most common is hypoxic treatment of the cells. This can be also be chemically induced by CoCl2 treatment as well. Our recommended protocols and lysate preparations can be found in our white paper here: http://www.novusbio.com/whitepapers/cellular-response-hypoxia.html.
10. For paraffin sections, do you recommend the antigen retrieval?
    • Unless specially noted in the protocol section we always recommend using citrate buffer pH 6 for antigen retrieval. 

Bioinformatics

• Gene Symbol: EPAS1