HIF-1 alpha Antibody

Knockdown Validated: HIF-1 alpha Antibody [NB100-449] - Morphological alterations induced by DMOG are HIF-1alpha dependent. glEND.2 cells were stably transfected with shGFP, shHIF-1alpha or shHIF-2alpha. The clones were cultured under normoxic (N) or hypoxic (H, 1% O2) conditions, or treated with DMOG for 6 h. 30 ug protein of cellular protein were loaded for the detection of HIF-1alpha, and nuclear extracts (20 ug protein) were used for the detection of HIF-2alpha. Image collected and cropped by CiteAb from the following publication (//biosignaling.biomedcentral.com/articles/10.1186/1478-811X-11-80) licensed under a CC-BY licence.

Western Blot: HIF-1 alpha Antibody [NB100-449] - HIF-1 alpha induction on Caki-1 cell lysate using CoCl2. Image from verified customer review.

Simple Western: HIF-1 alpha Antibody [NB100-449] - Simple Western lane view shows a specific band for HIF-1 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-1 alpha Antibody [NB100-449] - Mouse Brain, Neurons 40X

Immunohistochemistry: HIF-1 alpha Antibody [NB100-449] - Detection of human HIF1-alpha by immunohistochemistry. Sample: FFPE section of renal cell carcinoma. Antibody: Affinity purified rabbit anti-HIF1-alpha antibody (NB110-58773). Detection: DAB

Western Blot: HIF-1 alpha Antibody [NB100-449] - Detection of Human HIF-1 alpha by Western Blot and Immunoprecipitation. Samples: Whole cell lysate (5, 15 and 50 ug for WB; 1 mg for IP, 20% of IP loaded) from HeLa cells that were either treated with cobalt chloride (+; 200 uM) or mock treated (-). Antibodies: Affinity purified rabbit anti-HIF1 alpha antibody used for WB at 0.1 ug/mL(A) and 1 ug/mL (B) and used for IP at 3 ug/mg lysate. HIF-1 alpha was also immunoprecipitated by a previous lot of this antibody. Detection: Chemiluminescence with exposure times of 30 seconds (A) and 10 seconds (B).

Western Blot: HIF-1 alpha Antibody [NB100-449] - 3T3-L1 mouse embryonic fibroblast adipose-like cell line. Antibody at 1:2000. WB image submitted by a verified customer review.

Immunocytochemistry/Immunofluorescence: HIF-1 alpha Antibody [NB100-449] - Murine primary bone marrow derived macrophages stained with HIF1-alpha antibody (red). Nuclei were counterstained with DAPI (blue). Image from verified customer review.

Immunohistochemistry-Paraffin: HIF-1 alpha Antibody [NB100-449] - Analysis of microvascular density and HIF-1alpha activity. Low, homogenous expression of HIF-1alpha was detected in the tumors of NNK treated mice (40X, scale bar = 25 um). Image collected and cropped by CiteAb from the following publication (//molecular-cancer.biomedcentral.com/articles/10.1186/1476-4598-11-4), licensed under a CC-BY licence.

Flow Cytometry: HIF-1 alpha Antibody [NB100-449] - HeLa cells were treated for 15 hrs with 200 uM CoCl2, fixed in PFA, and permeabilized in 90% MeOH. 1 X 10^6 cells were stained with 0.125 ug anti-HIF-1 alpha and secondary FITC-conjugated goat anti-rabbit (in a 150 uL reaction). Black: treated, anti-KLH control IgG; Red: untreated, anti-HIF-1 alpha; Blue: treated, anti-HIF-1 alpha.

Western Blot: HIF-1 alpha Antibody [NB100-449] - Analysis of HIF-1 alpha in human myeloma cell lysate using anti-HIF-1 alpha. Cells were untreated or treated with IGF-1, IL-6 or CoCl2. Image from verified customer review.

Western Blot: HIF-1 alpha Antibody [NB100-449] - Detection of HIF-1 alpha in a hypoxic sample. Lane 1: CoCl2 treated Cos-7 nuclear extract (hypoxic). Lane 2: Untreated Cos-7 nuclear extract (normoxic).

Western Blot: HIF-1 alpha Antibody [NB100-449] - Detection of mouse HIF-1 alpha on hypoxia treated MEFs

Western Blot: HIF-1 alpha Antibody [NB100-449] - Samples: Whole cell lysate (5, 15 and 50 ug) from HeLa cells that were treated with cobalt chloride (+; 200 uM) or mock treated (-). Antibodies: Affinity purified rabbit anti-HIF1 alpha antibody NB100-449 used for WB at 0.1 ug/ml. Detection: Chemiluminescence with exposure times of 30 seconds.

Western Blot: HIF-1 alpha Antibody [NB100-449] - BMDM were seeded at 0.5x10^6 overnight. Cells were treated with 10 ng/ml LPS for 24 hrs. Image from verified customer review.

Western Blot: HIF-1 alpha Antibody [NB100-449] - Analysis of total protein using anti-HIF-1 alpha, -CAIX, -ISCU antibodies and VDACs antibody (PMID: 29596470).

Western Blot: HIF-1 alpha Antibody [NB100-449] - Analysis of total protein using anti-HIF-1 alpha, -ISCU, -FXN antibodies. CoCl2 was used to treat or not HeLa cells for 2 days (PMID: 29596470).

Western Blot: HIF-1 alpha Antibody [NB100-449] - Detection of human HIF1 alpha by western blot. Samples: Whole cell lysate (15 and 50 ug) from HEK293T cells that were either treated with 200 uM cobalt chloride (+) or mock treated (-). Antibodies: Affinity purified rabbit anti-HIF1 alpha antibody NB100-449 used for WB at 0.1 ug/ml. Detection: Chemiluminescence with exposure times of 3 minutes.

Immunocytochemistry/Immunofluorescence: HIF-1 alpha Antibody [NB100-449] - Formaldehyde-fixed asynchronous HeLa cells.

Immunoprecipitation: HIF-1 alpha Antibody [NB100-449] - HIF-1 alpha analysis in HEK293 cells. Image from verified customer review.

Immunoprecipitation: HIF-1 alpha Antibody [NB100-449] - Detection of human HIF1 alpha by western blot of immunoprecipitates. Samples: Whole cell lysate (1 mg for IP, 20% of IP loaded) from HEK293T cells that were treated with 200 uM cobalt chloride. Antibodies: Affinity purified rabbit anti-HIF1 alpha antibody NB100-449 used for WB at 1 ug/ml and for IP at 6 ug/mg lysate. Detection: Chemiluminescence with exposure times of 3 minutes.

• Reactivity: Hu, Mu, Rt, Ca, Ch, Gt, Pm, Pm, Bv
• Applications: WB, Simple Western, ChIP, ELISA, Flow, Flow-IC, ICC/IF, IHC, IHC-Fr, IHC-P, IP, KD, KO
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 0.2 mg/ml

HIF-1 alpha Antibody Summary

• Immunogen: The immunogen recognized by this HIF-1 alpha Antibody maps to a region between residues 775 and the C-terminus (residue 826) of human hypoxia-inducible factor 1 [Uniprot# Q16665].
• Predicted Species: Bovine (100%). Backed by our 100% Guarantee.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: HIF1A
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Western Blot 1:2000 - 1:10000
  • Simple Western 1:200
  • Chromatin Immunoprecipitation 1:10 - 1:500
  • ELISA 1:100-1:2000
  • Flow Cytometry 0.125 ug per 1 million cells in a 150 mcl reaction
  • Immunocytochemistry/Immunofluorescence 1:10-1:500
  • Immunohistochemistry 1:100 - 1:500
  • Immunohistochemistry-Frozen 1:50-1:200
  • Immunohistochemistry-Paraffin 1:50-1:200
  • Immunoprecipitation 2-5 ug/mg lysate
  • Flow (Intracellular)
  • Knockdown Validated
  • Knockout Validated
• Application Notes: ChIP usage was reported in scientific literature (PMID: 25557133). In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. For IHC-P, Tris-EDTA pH 9.0 buffer is recommended for the heat induced epitope retrieval. ELISA (PMID: 17556599 and 16966370). Knockout data (PMID: 31793879). Use in Flow-intracellular reported in scientific literature (PMID:31722203).
• Theoretical MW: 93 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:

  Human Brain Cerebral Cortex Whole Tissue Lysate (Adult Whole Normal)
  Human Pancreas Whole Tissue Lysate (Adult Whole Tumor)
  COS-7 Nuclear Hypoxic Induced Cell Lysate
  HeLa CoCl2 treated/Untreated Cell Lysate
  HepG2 CoCl2 treated/Untreated Cell Lysate
  HeLa Hypoxic / Normoxic Cell Lysate
  HepG2 Hypoxic / Normoxic Cell Lysate
  HIF-1 alpha Knockout HeLa Cell Lysate

Reactivity Notes

Monkey (COS-7) and Rat reactivities were reported in customer review feedback. Detection of HIF1 alpha in both Mouse and Human tissue by IHC. Chicken reactivity was reported in scientific literature (PMID: 25632022). Canine reactivity reported in scientific literature (PMID: 28701694). Reactivity with Goat is reported in PMID: 21599540.

Packaging, Storage & Formulations

• Storage: Store at 4C. Do not freeze.
• Buffer: TBS, 0.1% BSA
• Preservative: 0.09% Sodium Azide
• Concentration: 0.2 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for HIF-1 alpha Antibody

• AINT
• anti-HIF-1 alpha
• anti-HIF1A
• ARNT interacting protein
• ARNT-interacting protein
• Basic-helix-loop-helix-PAS protein MOP1
• BHLHE78
• Class E basic helix-loop-helix protein 78
• HIF 1A
• HIF1 alpha
• HIF-1 alpha
• HIF1
• HIF1A
• HIF-1a
• HIF-1alpha
• HIF-1-alpha
• HIF1-alpha
• hypoxia inducible factor 1 alpha subunit, hypoxia inducible factor 1 subunit alpha
• hypoxia inducible factor 1, alpha subunit (basic helix-loop-helix transcription factor)
• hypoxia-inducible factor 1-alpha
• Member of PAS protein 1
• member of PAS superfamily 1
• MOP1
• PAS domain-containing protein 8
• PASD8

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 (predicted molecular weight 93kDa), 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. In its active form under hypoxic conditions, HIF-1 is stabilized by the formation of a heterodimer of HIF-1 alpha and ARNT/HIF-1 beta subunits. Nuclear HIF-1 engages p300/CBP for binding to hypoxic response elements (HREs). This process induces transcription and regulation of genes including EPO, VEGF, iNOS2, ANGPT1 and OCT4 (4,5).

Under normoxic conditions, the HIF-1 alpha subunit is rapidly targeted and degraded by the ubiquitin proteasome system. This process is mediated by prolyl hydroxylase domain enzymes (PHDs), which catalyze the hydroxylation of key proline residues (Pro-402 and Pro-564) within the oxygen-dependent degradation domain of HIF-1 alpha. Once hydroxylated, HIF-1 alpha binds the von Hippel-Lindau tumor suppressor protein (pVHL) for subsequent ubiquitination and proteasomal degradation (4). pVHL dependent regulation of HIF-1 alpha plays a role in normal physiology and disease states. Regulation of HIF-1 alpha by pVHL is critical for the suppressive function of Foxp3+ regulatory Tcells (6). Repression of pVHL expression in chronic lymphocytic leukemia (CLL) B cells leads to HIF-1 alpha stabilization and increased VEGF secretion (7).

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. 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

5. Dengler, V. L., Galbraith, M. D., & Espinosa, J. M. (2014). Transcriptional regulation by hypoxia inducible factors. Critical Reviews in Biochemistry and Molecular Biology. https://doi.org/10.3109/10409238.2013.838205

6. Lee, J. H., Elly, C., Park, Y., & Liu, Y. C. (2015). E3Ubiquitin Ligase VHL Regulates Hypoxia-Inducible Factor-1 alpha to Maintain Regulatory T Cell Stability and Suppressive Capacity. Immunity. https://doi.org/10.1016/j.immuni.2015.05.016

7. Ghosh, A. K., Shanafelt, T. D., Cimmino, A., Taccioli, C., Volinia, S., Liu, C. G.,... Kay, N. E. (2009). Aberrant regulation of pVHL levels by microRNA promotes the HIF/VEGF axis in CLL B cells. Blood. https://doi.org/10.1182/blood-2008-10-185686

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-1 alpha Antibody (NB100-449)(231)

Publications using NB100-449

• Ghosh P, Guo Y, Ashrafi A et al. Oxygen-enhanced optoacoustic tomography reveals the effectiveness of targeting heme and oxidative phosphorylation at normalizing tumor vascular oxygenation Cancer Res. Jun 16 2020 [PMID: 32546631] (IHC, Human)
• Hu L, Chen X, Qiu S et al. Intra-Pancreatic Insulin Nourishes Cancer Cells: Do Insulin-Receptor Antagonists such as PGG and EGCG Play a Role? Am. J. Chin. Med. May 28 2020 [PMID: 32468825]
• Rawat S Maturation of Cardiac Energy Metabolism in the Normal and Hypertrophied Newborn Heart Thesis Jan 1 2019 (WB, Human)
• Kim KY, Kim YR, Choi KW et al. Downregulated miR-18b-5p triggers apoptosis by inhibition of calcium signaling and neuronal cell differentiation in transgenic SOD1 (G93A) mice and SOD1 (G17S and G86S) ALS patients Transl Neurodegener Jul 1 2020 [PMID: 32605607]
• Leinhos L Myofibroblast differentiation in hypoxia: a novel role for ArhGAP29 Thesis (WB)
• van Dam TP DNA polymerase beta prevents AID-instigated mutagenic non-canonical mismatch DNA repair bioRxiv Jan 1 2020 (IP, Mouse)
• Sil S, Hu G, Liao K et al. HIV-1 Tat-mediated astrocytic amyloidosis involves the HIF-1 alpha/lncRNA BACE1-AS axis PLoS Biol. May 26 2020 [PMID: 32453744] (ICC/IF, Human)
• Doigneaux C, Pedley AM, Mistry IN et al. Hypoxia Drives the Assembly of the Multi-Enzyme Purinosome Complex J. Biol. Chem. May 21 2020 [PMID: 32439803] (WB, Human)
• Sola-Penna M, PaixAo LP, Branco JR, et al. Serotonin activates glycolysis and mitochondria biogenesis in human breast cancer cells through activation of the Jak1/STAT3/ERK1/2 and adenylate cyclase/PKA, respectively Br. J. Cancer Dec 10 2019 [PMID: 31819176] (WB, Human)
• Di Gioia M, Spreafico R, Springstead JR et al. Endogenous oxidized phospholipids reprogram cellular metabolism and boost hyperinflammation Nat. Immunol. Nov 25 2019 [PMID: 31768073] (WB, Mouse)
• Gupta SS, Sharp R, Hofferek C et al. NIX-Mediated Mitophagy Promotes Effector Memory Formation in Antigen-Specific CD8+ T Cells Cell Rep Nov 12 2019 [PMID: 31722203] (Flow-IC, Mouse)
• Marrocco A Alterations of CCSP expression and macrophages metabolism in the development of silica-induced pulmonary inflammation and fibrosis Thesis Jan 1 2020 (ICC/IF, Mouse)
• Yamazaki H, Tanaka T, Mie K et al. Assessment of postoperative adjuvant treatment using toceranib phosphate against adenocarcinoma in dogs J. Vet. Intern. Med. Apr 8 2020 [PMID: 32267594] (IHC-P, Canine)
• Das Gupta K, Shakespear MR, Curson JEB et al. Class IIa Histone Deacetylases Drive Toll-like Receptor-Inducible Glycolysis and Macrophage Inflammatory Responses via Pyruvate Kinase M2 Cell Rep Feb 25 2020 [PMID: 32101747] (WB, Human)
• Sugahara M, Tanaka S, Tanaka T et al. Prolyl Hydroxylase Domain Inhibitor Protects against Metabolic Disorders and Associated Kidney Disease in Obese Type 2 Diabetic Mice J. Am. Soc. Nephrol. Jan 29 2020 [PMID: 31996409]
• Yambire KF, Rostosky C, Watanabe T Impaired lysosomal acidification triggers iron deficiency and inflammation in vivo Elife Dec 3 2019 [PMID: 31793879] (WB, KO, Mouse)
• 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]
• Lin HY, Chou PC, Chen YH et al. Dietary Supplementation of 25-Hydroxycholecalciferol Improves Livability in Broiler Breeder Hens-Amelioration of Cardiac Pathogenesis and Hepatopathology Animals (Basel) 2019 Oct 08 [PMID: 31597394] (WB, Chicken)
• Nasyrov E, Nolan KA, Wenger RH et al. The neuronal oxygen-sensing pathway controls postnatal vascularization of the murine brain FASEB J. Aug 30 2019 [PMID: 31469589]
• Bailey JD, Diotallevi M, Nicol T, et al. Nitric Oxide Modulates Metabolic Remodeling in Inflammatory Macrophages through TCA Cycle Regulation and Itaconate Accumulation Cell Rep 2019 Jul 2 [PMID: 31269442] (WB)
• Bahrami B, Shen W, Zhu L et al. Effects of VEGF inhibitors on human retinal pigment epithelium under high glucose and hypoxia Clin. Experiment. Ophthalmol. Jul 2 2019 [PMID: 31265210]
• Baik SH, Kang S, Lee W et al. A Breakdown in Metabolic Reprogramming Causes Microglia Dysfunction in Alzheimer's Disease Cell Metab. Jun 24 2019 [PMID: 31257151]
• Chowdhury D, Alrefai H, Landero Figueroa JA et al. Metallothionein 3 Controls the Phenotype and Metabolic Programming of Alternatively Activated Macrophages Cell Rep Jun 25 2019 [PMID: 31242420] (ICC/IF, Mouse)
• Chen Y, Zhao B, Zhu Y et al. HIF-1-VEGF-Notch mediates angiogenesis in temporomandibular joint osteoarthritis Am J Transl Res. May 15 2019 [PMID: 31217867] (IHC, WB, Rat)
• Bechmann N, Poser I, Seifert V et al. Dendritic Cell-Derived TSLP Negatively Regulates HIF-1 alpha and IL-1 beta During Dectin-1 Signaling Front Immunol May 8 2019 [PMID: 31139177] (WB, Human)
• Elder MJ, Webster SJ, Fitzmaurice TJ et al. Digoxin Attenuates Receptor Activation of NF-kappa B Ligand-Induced Osteoclastogenesis in Macrophages J. Vasc. Res. May 14 2019 [PMID: 31085912] (WB, Mouse)
• Duan LJ, Fong GH. Developmental vascular pruning in neonatal mouse retinas is programmed in the astrocytic oxygen sensing mechanism Development Mar 25 2019 [PMID: 30910827] (WB, Mouse)
• Grigelioniene G, Suzuki HI, Taylan F et al. Gain-of-function mutation of microRNA-140 in human skeletal dysplasia Nat. Med. Feb 25 2019 [PMID: 30804514] (WB, Mouse)
• Li HS, Zhou YN, Li L et al. HIF-1a protects against oxidative stress by directly targeting mitochondria. Redox Biol Jan 14 2019 [PMID: 30686776] (IHC-P, IP, Human)
• Krueger K, Catanese L, Sciesielski LK et al. Deletion of an intronic HIF-2a binding site suppresses hypoxia-induced WT1 expression. Biochim Biophys Acta Gene Regul Mech. Nov 20 2018 [PMID: 30468780] (ChIP, Human)
• Bae YH, Joo H, Bae J et al. Brain injury induces HIF-1a-dependent transcriptional activation of LRRK2 that exacerbates brain damage. Cell Death Dis. Nov 12 2018 [PMID: 30420654] (ChIP, Mouse)
• Valencia-Cervantes J, Huerta-Yepez S, Aquino-Jarquin G et al. Hypoxia increases chemoresistance in human medulloblastoma DAOY cells via hypoxia‑inducible factor 1a‑mediated downregulation of the CYP2B6, CYP3A4 and CYP3A5 enzymes and inhibition of cell proliferation. Oncol. Rep. Oct 12 2018 [PMID: 30320358] (Human)
• Fauzia E, Barbhuyan Tk, Shrivastava Ak et al. Chick Embryo: A Preclinical Model for Understanding Ischemia-Reperfusion Mechanism. Front Pharmacol. Sep 21 2018 [PMID: 30298003] (WB, Chicken)
• Mouly L, Mamouni K, Gence R et al. PARP-1-dependent RND1 transcription induced by topoisomerase I cleavage complexes confers cellular resistance to camptothecin. Cell Death Dis Sep 12 2018 [PMID: 30209297] (WB, Human)
• Koenis DS, Medzikovic L, van Loenen PB et al. Nuclear Receptor Nur77 Limits the Macrophage Inflammatory Response through Transcriptional Reprogramming of Mitochondrial Metabolism Cell Rep Aug 21 2018 [PMID: 30134173] (WB, Mouse)
• Koenis D, Medzikovic L, Vos M et al. Nur77 variants solely comprising the amino-terminal domain activate hypoxia-inducible factor-1a and affect bone marrow homeostasis in mouse and man J. Biol. Chem. Aug 15 2018 [PMID: 30111591] (WB, Mouse)
• kim YE, Lee M, Gu H et al. HIF-1a activation in myeloid cells accelerates dextran sodium sulfate-induced colitis progression in mice. Dis Model Mech. Jul 30 2018 [PMID: 29967068] (IHC-Fr, Mouse)
• Li B, Zhu Y, Sun Q et al. Reversal of the Warburg effect with DCA in PDGF-treated human PASMC is potentiated by pyruvate dehydrogenase kinase-1 inhibition mediated through blocking Akt/GSK-3 beta signalling Int J Mol Med. 2018 Jun 26 [PMID: 29956736] (WB, Human)
• Mekki MS, Mougel A, Vinchent A et al. Hypoxia leads to decreased autophosphorylation of the MET receptor but promotes its resistance to tyrosine kinase inhibitors Oncotarget Jun 5 2018 [PMID: 29930749] (WB, Human)
• Hamann I, Krys D, Glubrecht D et al. Expression and function of hexose transporters GLUT1, GLUT2, and GLUT5 in breast cancer-effects of hypoxia The FASEB Journal 2018 Apr 13 [PMID: 29913554] (IHC, Human)
• Niland S, Komljenovic D, Macas J et al. Rhodocetin-aB selectively breaks the endothelial barrier of the tumor vasculature in HT1080 fibrosarcoma and A431 epidermoid carcinoma tumor models Oncotarget Apr 27 2018 [PMID: 29854288] (WB)
• Fukushima A, Zhang L, Huqi A et al. Acetylation contributes to hypertrophy-caused maturational delay of cardiac energy metabolism JCI Insight May 17 2018 [PMID: 29769443] (WB, Human)
• Ferecatu I, Canal F, Fabbri L et al. Dysfunction in the mitochondrial Fe-S assembly machinery leads to formation of the chemoresistant truncated VDAC1 isoform without HIF-1a activation. PLoS ONE Mar 29 2018 [PMID: 29596470] (WB, Human)
• Cho JH, Kim GY, Mansfield BC, Chou JY. Hepatic glucose-6-phosphatase-A deficiency leads to metabolic reprogramming in glycogen storage disease type Ia. Biochem. Biophys. Res. Commun. Apr 15 2018 [PMID: 29545180] (Mouse)
• Bok S, Kim YE, Woo Y et al. Hypoxia-inducible factor-1a regulates microglial functions affecting neuronal survival in the acute phase of ischemic stroke in mice Oncotarget 2017 Dec 19 [PMID: 29340071] (WB, IHC-Fr, Mouse)
• Geoghegan F, Buckland RJ, Rogers E et al. Bioenergetics of acquired cisplatin resistant H1299 non-small cell lung cancer and P31 mesothelioma cells Oncotarget. 2017 Oct 16 [PMID: 29212260] (Human)
• Almiron Bonnin DA, Havrda MC, Lee MC et al. Secretion-mediated STAT3 activation promotes self-renewal of glioma stem-like cells during hypoxia Oncogene. 2017 Nov 20 [PMID: 29155422] (WB, Mouse)
• Palazon A, Tyrakis PA, Macias D et al. An HIF-1 alpha/VEGF-A Axis in Cytotoxic T Cells Regulates Tumor Progression Cancer Cell 2017 Nov 13 [PMID: 29136509] (WB, Mouse)
• Palsson-McDermott EM, Dyck L, Zastona Z et al. Pyruvate Kinase M2 Is Required for the Expression of the Immune Checkpoint PD-L1 in Immune Cells and Tumors. Front Immunol. 2017 Oct 30 [PMID: 29081778] (WB, Mouse)
• Lee S, Kim JH, Lee JH et al. Imaging Monitoring of Kupffer Cell Function and Hepatic Oxygen Saturation in Preneoplastic Changes During Cholangiocarcinogenesis Sci Rep. 2017 Oct 27 [PMID: 29079853] (WB, Mouse)
• Domingo-Gonzalez R, Das S, Griffiths KL et al. Interleukin-17 limits hypoxia-inducible factor 1a and development of hypoxic granulomas during tuberculosis. JCI Insight. 2017 Oct 05 [PMID: 28978810] (WB, Mouse)
  
  Details:
  HIF1 alpha antibody used to evaluate the effect of IL-17 on the shift to glycolysis and the development of hypoxic tuberculosis granulomas
• Goodwin J, Choi H, Hsieh MH et al. Targeting HIF-1a/PDK1 Axis by Dichloroacetate (DCA) Suppresses Bleomycin-induced Pulmonary Fibrosis Am. J. Respir. Cell Mol. Biol. 2017 Sep 15 [PMID: 28915065] (Mouse)
• Huang R, Hayashi Y, Yan X, et al. HIF1A is a critical downstream mediator for hemophagocytic lymphohistiocytosis. Haematologica 2017 Aug 31 [PMID: 28860338] (WB, Mouse)
• MacLauchlan SC, Calabro NE, Huang Y et al. HIF-1alpha represses the expression of the angiogenesis inhibitor thrombospondin-2 Matrix Biol. 2017 Aug 05 [PMID: 28789925] (WB, Mouse)
• Lee S, Kim JH, Lee JH et al. Non-invasive monitoring of the therapeutic response in sorafenib-treated hepatocellular carcinoma based on photoacoustic imaging. Eur Radiol 2017 Jul 27 [PMID: 28752217] (ICC/IF, Mouse)
• Elder MJ, Webster SJ, Chee R et al. B-Glucan Size Controls Dectin-1-Mediated Immune Responses in Human Dendritic Cells by Regulating IL-1B Production Front Immunol 2017 Jul 24 [PMID: 28736555] (WB, Human)
• Tavernier SJ, Vanlangenakker N, Vetters J et al. Opposing regulation and roles for PHD3 in lung dendritic cells and alveolar macrophages J. Leukoc. Biol. 2017 Jul 17 [PMID: 28716863] (WB)
• Zhang JT, Wang Y, Chen JJ et al. Defective CFTR leads to aberrant B-catenin activation and kidney fibrosis Sci Rep 2017 Jul 12 [PMID: 28701694] (WB, Canine)
• Papathanassiu AE, Ko JH, Imprialou M et al. BCAT1 controls metabolic reprogramming in activated human macrophages and is associated with inflammatory diseases Nat Commun. 2017 Jul 11 [PMID: 28699638] (WB, Mouse)
• Carlessi R, Chen Y, Rowlands J et al. GLP-1 receptor signalling promotes B-cell glucose metabolism via mTOR-dependent HIF-1alpha activation. Sci Rep. 2017 Jun 01 [PMID: 28572610] (WB, Mouse)
• Goodwin J, Neugent ML, Lee SY et al. The distinct metabolic phenotype of lung squamous cell carcinoma defines selective vulnerability to glycolytic inhibition. Nat Commun. 2017 May 26 [PMID: 28548087] (IHC-P, Human)
• Feigenson M, Eliseev RA, Jonason JH et al. PGE2 Receptor Subtype 1 (EP1) Regulates Mesenchymal Stromal Cell Osteogenic Differentiation by Modulating Cellular Energy Metabolism. J. Cell. Biochem. Apr 26 2017 [PMID: 28444901] (WB, Mouse)
• Kowalski GM, Kraakman MJ, Mason SA et al. Resolution of glucose intolerance in long-term high-fat, high-sucrose-fed mice. J. Endocrinol. 2017 Jun 01 [PMID: 28360081] (WB, Mouse)
• Evans Colin E, Palazon Asis, Sim Jingwei et al. Modelling pulmonary microthrombosis coupled to metastasis: distinct effects of thrombogenesis on tumorigenesis. Biology Open 2017 [PMID: 28302670] (IHC, Mouse)
• Shen W, Yau B, Lee SR et al. Effects of Ranibizumab and Aflibercept on Human Muller Cells and Photoreceptors under Stress Conditions. Int J Mol Sci. Mar 1 2017 [PMID: 28257068] (WB, Human)
• Gardner PJ, Liyanage SE, Cristante E et al. Hypoxia inducible factors are dispensable for myeloid cell migration into the inflamed mouse eye Sci Rep Jan 23 2017 [PMID: 28112274] (WB, Mouse)
• McGettrick AF, Corcoran SE, Barry PJ et al. Trypanosoma brucei metabolite indolepyruvate decreases HIF-1alpha and glycolysis in macrophages as a mechanism of innate immune evasion. Proc. Natl. Acad. Sci. U.S.A. Nov 29 2016 [PMID: 27856732]
• Wang Y, Wang H, Li J et al. Direct visualization of the phenotype of hypoxic tumor cells at single cell resolution in vivo using a new hypoxia probe. Intravital. Oct 28 2016 [PMID: 27790387] (Mouse, Human)
• Mills El, Kelly B, Logan A et al. Succinate Dehydrogenase Supports Metabolic Repurposing of Mitochondria to Drive Inflammatory Macrophages. Cell Sep 21 2016 [PMID: 27667687]
• Alonso J, Galan M, Marti-Pamies I et al. NOR-1/NR4A3 regulates the cellular inhibitor of apoptosis 2 (cIAP2) in vascular cells: role in the survival response to hypoxic stress. Sci Rep 2016 Sep 22 [PMID: 27654514] (WB)
• Turlejski T, Humoud I, Desai R et al. Immunohistochemical evidence of tissue hypoxia and astrogliosis in the rostral ventrolateral medulla of spontaneously hypertensive rats. Brain Res. Sep 8 2016 [PMID: 27616338] (IHC)
• Prakash H, Nadella V, Singh S, Schmitz-Winnenthal H. CD14/TLR4 priming potentially recalibrates and exerts anti-tumor efficacy in tumor associated macrophages in a mouse model of pancreatic carcinoma. Sci Rep. Aug 11 2016 [PMID: 27511884]
• Lampropoulou V, Sergushichev A, Bambouskova M et al. Itaconate Links Inhibition of Succinate Dehydrogenase with Macrophage Metabolic Remodeling and Regulation of Inflammation Cell Metab. 2016 Jul 12 [PMID: 27374498] (WB, Mouse)
• Mirzamohammadi F, Papaioannou G, Inloes JB et al. Polycomb repressive complex 2 regulates skeletal growth by suppressing Wnt and TGF-b signalling. Nat Commun Jun 22 2016 [PMID: 27329220] (WB, Mouse)
• Pei L, Solis G, Nguyen MT et al. Paracellular epithelial sodium transport maximizes energy efficiency in the kidney J. Clin. Invest Jul 1 2016 [PMID: 27214555] (WB, Mouse)
• Semba H, Takeda N, Isagawa T et al. HIF-1alpha-PDk1 axis-induced active glycolysis plays an essential role in macrophage migratory capacity. Nat Commun. May 18 2016 [PMID: 27189088] (WB, Mouse)
• Tu DG, Chang WW, Lin ST et al. Salmonella inhibits tumor angiogenesis by downregulation of vascular endothelial growth factor. Oncotarget. Jun 21 2016 [PMID: 27175584] (WB, Mouse)
• Kobayashi H, Liu Q, Binns TC et al. Distinct subpopulations of FOXD1 stroma-derived cells regulate renal erythropoietin. J. Clin. Invest. 2016 May 02 [PMID: 27088801] (WB, Mouse)
• Nusken E, Wohlfarth M, Lippach G et al. Reduced Perinatal Leptin Availability May Contribute to Adverse Metabolic Programming in a Rat Model of Uteroplacental Insufficiency. Endocrinology 2016 May [PMID: 27007072]
• Kurihara Toshihide, Westenskow Peter D, Gantner Marin L et al. Hypoxia-induced metabolic stress in retinal pigment epithelial cells is sufficient to induce photoreceptor degeneRation. ELife 2016 [PMID: 26978795] (IHC, Mouse)
• Shukla PK, Chaudhry KK, Mir H et al. Chronic ethanol feeding promotes azoxymethane and dextran sulfate sodium-induced colonic tumorigenesis potentially by enhancing mucosal inflammation BMC Cancer Mar 8 2016 [PMID: 26951793] (IHC-Fr, Mouse)
• Farsijani NM, Liu Q, Kobayashi H et al. Renal epithelium regulates erythropoiesis via HIF-dependent suppression of erythropoietin. J Clin Invest 2016 Apr 1 [PMID: 26927670]
• Chapple SJ, Keeley TP, Mastronicola D et al. Bach1 differentially regulates distinct Nrf2-dependent genes in human venous and coronary artery endothelial cells adapted to physiological oxygen levels. Free Radic. Biol. Med. 2015 Dec 15 [PMID: 26698668] (WB, Human)
• Shum LC, White NS, Mills BN et al. Energy Metabolism in Mesenchymal Stem Cells During Osteogenic Differentiation. Stem Cells Dev 2016 Jan 15 [PMID: 26487485]
• Konzack A, Jakupovic M, Kubaichuk K et al. Mitochondrial Dysfunction Due to Lack of Manganese Superoxide Dismutase Promotes Hepatocarcinogenesis. Antioxid Redox Signal 2015 Nov 10 [PMID: 26422659] (IHC)
• Lojk J, Mis K, Pirkmajer S, Pavlin M. siRNA delivery into cultured primary human myoblasts - optimization of electroporation parameters and theoretical analysis. Bioelectromagnetics 2015 Sep 21 [PMID: 26388450] (WB, Human)
• Niebler S, Angele P, Kujat R, Bosserhoff AK. Hypoxia-Inducible Factor 1 Is an Inductor of Transcription Factor Activating Protein 2 Epsilon Expression during Chondrogenic Differentiation. Biomed Res Int. 2015 Aug 14 [PMID: 26273614] (WB, Mouse)
  
  Details:
  HIF-1 alpha antibody was used for WB on lysates of murine multipotent mesenchymal stem cells (Figure 4a) and SW1353 cells treated or not with siRNA against HIF1 alpha (Figure 4c).
• Miao H, Chen L, Hao L et al. Stearic acid induces proinflammatory cytokine production partly through activation of lactate-HIF1alpha pathway in chondrocytes. Sci Rep. 2015 Aug 14 [PMID: 26271607] (WB, Mouse)
• Le Moan N, Baeten KM, Rafalski VA et al. Hypoxia Inducible Factor-1 in Astrocytes and/or Myeloid Cells Is Not Required for the Development of Autoimmune Demyelinating Disease eNeuro 2015 Mar-Apr [PMID: 26213713] (IHC)
• Garhyan J, Bhuyan S, Pulu I et al. Preclinical and Clinical Evidence of Mycobacterium tuberculosis Persistence in the Hypoxic Niche of Bone Marrow Mesenchymal Stem Cells after Therapy. Am J Pathol 2015 Jul [PMID: 26066709]
• Fan D, Coughlin LA, Neubauer MM et al. Activation of HIF-1alpha and LL-37 by commensal bacteria inhibits Candida albicans colonization. Nat Med 2015 Jul [PMID: 26053625]
• Kambayashi S, Igase M, Kobayashi K et al. Hypoxia inducible factor 1alpha expression and effects of its inhibitors in canine lymphoma. J Vet Med Sci 2015 Nov [PMID: 26050843] (WB)
• Qiao S, Dennis M, Song X et al. A REDD1/TXNIP pro-oxidant complex regulates ATG4B activity to control stress-induced autophagy and sustain exercise capacity Nat Commun 2015 Apr 28 [PMID: 25916556] (WB, Human)
• Hribar K C, Finlay D, Ma X. et al. et al. Nonlinear 3D projection printing of concave hydrogel microstructures for long-term multicellular spheroid and embryoid body culture. Lab on a Chip 2015 [PMID: 25900329] (IHC, Human)
• Forsberg EA, Botusan I, Wang J et al. Carnosine decreases IGFBP1 production in db/db mice through suppression of HIF-1 J. Endocrinol. 2015 Apr 13 [PMID: 25869614] (WB, Mouse)
• 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)
• Henique C, Mansouri A, Vavrova E et al. Increasing mitochondrial muscle fatty acid oxidation induces skeletal muscle remodeling toward an oxidative phenotype FASEB J. 2015 Feb 23 [PMID: 25713059] (WB, Mouse)
• 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)
• Teranishi Y, Matsubara T, Krausz KW et al. Involvement of hepatic stellate cell cytoglobin in acute hepatocyte damage through the regulation of CYP2E1-mediated xenobiotic metabolism Lab. Invest. 2015 Feb 16 [PMID: 25686096] (WB, Mouse)
• Nusken E, Herrmann Y, Wohlfarth M et al. Strong hypoxia reduces leptin synthesis in purified primary human trophoblasts Placenta. 2015 Feb 03 [PMID: 25684657] (WB, Human)
• Robin E, Marcillac F, Raddatz E. A hypoxic episode during cardiogenesis down-regulates the adenosinergic system and alters the myocardial anoxic tolerance. Am. J. Physiol. Regul. Integr. Comp. Physiol. 2015 Jan 28 [PMID: 25632022] (WB, Chicken)
• Chang E, Paterno J, Duscher D et al. Exercise induces stromal cell-derived factor-1alpha-mediated release of endothelial progenitor cells with increased vasculogenic function Plast. Reconstr. Surg. 2015 Feb 01 [PMID: 25626819] (WB, Human)
• Palsson-McDermott EM, Curtis AM, Goel G et al. Pyruvate Kinase M2 Regulates Hif-1alpha Activity and IL-1beta Induction and Is a Critical Determinant of the Warburg Effect in LPS-Activated Macrophages Cell Metab. 2015 Jan 06 [PMID: 25565206] (WB, Mouse)
• 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)
• Mai J, Wang F, Qiu Q et al. Tachycardia pacing induces myocardial neovascularization and mobilizes circulating endothelial progenitor cells partly via SDF-1 pathway in canines. Heart Vessels 2016 Feb [PMID: 25491934]
• Kong Ly, Wei J, Haider As et al. Therapeutic targets in subependymoma. J. neuroimmunol. 2014 Oct 31 [PMID: 25465288] (WB, Human)
• Appel S, Turnwald Em, Ankerne J et al. Hypoxia-Mediated Soluble Fms-Like Tyrosine Kinase 1 Increase is not Attenuated in Interleukin 6-Deficient Mice. Reprod Sci. 2014 Nov 18 [PMID: 25415335]
• Kim HR, Kim JH, Choi EJ et al. Hyperoxygenation Attenuated a Murine Model of Atopic Dermatitis through Raising Skin Level of ROS. PLoS ONE. 2014 Oct 03 [PMID: 25275529] (IHC-P, Mouse)
  
  Details:
  HIF1 alpha antibody used for IHC-P on the AD lesions from hyperbaric oxygen therapy (HBOT) or applying an oxygen-carrying chemical, perfluorodecalin (PFD) treated murine model of AD that was developed by repeated application of a chemical irritant (1% 2,4-dinitrochlorobenzene) and house dust mite (Dermatophagoide farinae) extract on one ear of BALB/c mice. Figure 6 shows a decreased HIF1 alpha expression in the AD lesions of HBOT or PFD treated mice.
• Kim M, Neinast MD, Frank AP et al. ERalpha upregulates Phd3 to ameliorate HIF-1 induced fibrosis and inflammation in adipose tissue Mol Metab. 2014 Sep 01 [PMID: 25161887] (IP, WB, Mouse)
  
  Details:
  HIF-1 alpha antibody used for IP and WB applications in HIF-1 alpha ubiquitination assay on NIH3T3 cells that were transfected with expression vectors for pcDNA-mHIF1a, myc-Ub1, pFlag-ERa and shPHD3 (Figure 2D).
• Roychowdhury S, Chiang DJ, McMullen MR et al. Moderate, chronic ethanol feeding exacerbates carbon-tetrachloride-induced hepatic fibrosis via hepatocyte-specific hypoxia inducible factor 1a. Pharmacol Res Perspect 2014 Oct 01 [PMID: 25089199] (IHC-Fr, Mouse)
  
  Details:
  HIF1a/HIF-1 alpha antibody used for IHC-Fr on frozen liver sections of pair- or ethanol-fed C57BL/6J wild-type mice (Figure 1B) and hepatocyte-specific HIF1a-deficint mice /delta HepHIF1a-/- on C57BL/6 background fed ethanol or not + CCl4 injections 2X/week for 5 weeks (Figure S1).
• Appel S, Ankerne J, Appel J et al. CNN3 Regulates Trophoblast Invasion and Is Upregulated by Hypoxia in BeWo Cells. PLoS ONE 2014 Jul 23 [PMID: 25050546] (WB, Human)
  
  Details:
  HIF-1 alpha antibody used for WB at 1:1000 dilution as hypoxia marker in BeWo cells grown under normoxic or hypoxic conditions for 24 (Figure 5A) as well as serum starved BeWo cells treated with PBS or 200 uM CoCl2 for 6 h (Figure 5B).
• Colegio OR, Chu NQ, Szabo AL et al. Functional polarization of tumour-associated macrophages by tumour-derived lactic acid. Nature 2014 Jul 13 [PMID: 25043024] (WB, Mouse)
  
  Details:
  HIF1 alpha antibody used for WB on mouse's macrophage cells grown under normoxia /hypoxia and on cells stimulated with control DMEM or Lewis lung carcinoma tumor conditioned medium (Extended Data Figure 2a).
• Ciavardelli D, Rossi C, Barcaroli D et al. Breast cancer stem cells rely on fermentative glycolysis and are sensitive to 2-deoxyglucose treatment. Cell Death Dis 2014 Jul 18 [PMID: 25032859] (WB, Human)
  
  Details:
  HIF1 alpha antibody used for WB on human breast cancer tissues derived breast cancer stem cells (BCSCs) and SDAC, spheroid-derived adherent cells (Figure 2d).
• O?reilly VC, Floro KL, Shi H et al. Gene-environment interaction demonstrates the vulnerability of the embryonic heart. Dev. Biol. 3/25/2014 [PMID: 24657234] (IHC-P, IHC-Fr, WB, Mouse)
• 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)
• Duan LJ, Takeda K, Fong GH. Hypoxia inducible factor-2 alpha regulates the development of retinal astrocytic network by maintaining adequate supply of astrocyte progenitors. PLoS ONE 2014 Jan 29 [PMID: 24475033] (WB, Mouse)
• Reischl S, Li L, Walkinshaw G et al. Inhibition of HIF prolyl-4-hydroxylases by FG-4497 reduces brain tissue injury and edema formation during ischemic stroke. PLoS ONE 2014 Jan 10 [PMID: 24409307] (WB, Mouse)
• Rosafio K, Pellerin L. Oxygen tension controls the expression of the monocarboxylate transporter MCT4 in cultured mouse cortical astrocytes via a hypoxia-inducible factor-1alpha-mediated transcriptional regulation. Glia 2013 Dec 21 [PMID: 24375723] (WB, Mouse)
• Ouyang X, Ghani A, Malik A et al. Adenosine is required for sustained inflammasome activation via the A2A receptor and the HIF-1alpha pathway. Nat Commun 2013 Dec 19 [PMID: 24352507] (WB, Mouse)
• Zhu Y, Zhang L, Gidday JM. Role of hypoxia-inducible factor-1alpha in preconditioning-induced protection of retinal ganglion cells in glaucoma. Mol Vis. 2013 Nov 23 [PMID: 24319330] (IHC-P, ICC/IF, Mouse)
• Weidemann A, Breyer J, Rehm M et al. HIF-1alpha activation results in actin cytoskeleton reorganization and modulation of Rac-1 signaling in endothelial cells. Cell Commun Signal. 2013 Oct 21 [PMID: 24144209] (WB, Mouse)
• Doedens AL, Phan AT, Stradner MH et al. Hypoxia-inducible factors enhance the effector responses of CD8+ T cells to persistent antigen. Nat Immunol. 2013 Nov [PMID: 24076634] (WB, Mouse)
• Zhao H, Luo X, Zhou Z et al. Early treatment with xenon protects against the cold ischemia associated with chronic allograft nephropathy in rats. Kidney Int. 2013 Sep 11 [PMID: 24025645] (IHC-P, Rat)
• Clinkenbeard EL, Farrow EG, Summers LJ et al. Neonatal iron deficiency causes abnormal phosphate metabolism by elevating FGF23 in normal and ADHR mice. J Bone Miner Res 2013 Jul 22 [PMID: 23873717]
• Sugimoto M, Cutler A, Shen B et al. Inhibition of EGF Signaling Protects the Diabetic Retina from Insulin-Induced Vascular Leakage. Am J Pathol 2013 Jul 3 [PMID: 23831329] (WB, Human)
• Miranda E, Nordgren IK, Male AL et al. A Cyclic Peptide Inhibitor of HIF-1 Heterodimerization That Inhibits Hypoxia Signaling in Cancer Cells. J Am Chem Soc 2013 Jul 17 [PMID: 23796364] (Human)
• Shum LC. Mitochondrial Metabolism in Bone Physiology and Pathology Thesis 2018 (WB, Human)
• Brackmann FA, Link AS, Jung S et al. Activin A regulation under global hypoxia in developing mouse brain. Brain Res 2013 Jul 31 [PMID: 23916668] (WB, Mouse)
• Piao Y, Liang J, Holmes L et al. Acquired Resistance to Anti-VEGF Therapy in Glioblastoma Is Associated with a Mesenchymal Transition. Clin Cancer Res 2013 Aug 15 [PMID: 23804423] (IHC-P, Human)
• Oh JH, Oh J, Togloom A et al. Effects of Ginkgo biloba Extract on Cultured Human Retinal Pigment Epithelial Cells under Chemical Hypoxia. Curr Eye Res 2013 Jun 21 [PMID: 23790153] (ICC/IF, Human)
• Danza G, Di Serio C, Ambrosio MR et al. Notch3 is activated by chronic hypoxia and contributes to the progression of human prostate cancer. Int J Cancer 2013 Jun 1 [PMID: 23729168] (WB, Human)
• Pawlak EA, Geor RJ, Watts MR et al. Regulation of hypoxia inducible factor-1alpha and related genes in equine digital lamellae and in cultured keratinocytes. Equine Vet J 2013 Apr 18 [PMID: 23663159]
• Balamurugan K, Sharan S, Klarmann KD et al. FBXW7alpha attenuates inflammatory signalling by downregulating C/EBPdelta and its target gene Tlr4. Nat Commun 2013 Apr 9 [PMID: 23575666] (WB, Human, Mouse)
• Tannahill GM, Curtis AM, Adamik J et al. Succinate is an inflammatory signal that induces IL-1beta through HIF-1alpha Nature 2013 Mar 24 [PMID: 23535595] (WB, Mouse)
• Baek Kwang Je, Cho Jae Youn, Rosenthal Peter et al. Hypoxia potentiates allergen induction of HIF-1a, chemokines, airway inflammation, TGF-B1, and airway remodeling in a mouse model. Clinical Immunology (Orlando, Fla.) 2013 [PMID: 23499929] (IHC, Mouse)
• Kocemba KA, van Andel H, de Haan-Kramer A et al. The hypoxia target adrenomedullin is aberrantly expressed in multiple myeloma and promotes angiogenesis Leukemia 2013 Mar 12 [PMID: 23478664] (WB, Human)
• Longati P, Jia X, Eimer J et al. 3D pancreatic carcinoma spheroids induce a matrix-rich, chemoresistant phenotype offering a better model for drug testing BMC Cancer 2013 Feb 27 [PMID: 23446043] (WB, Human)
• Huang C, Hales BF. Effects of exposure to a DNA damaging agent on the hypoxia inducible factors in organogenesis stage mouse limbs PLoS One 2012 [PMID: 23251655] (IHC, Mouse)
• Sun K, Halberg N, Khan M et al. Selective Inhibition of HIF1alpha Ameliorates Adipose Tissue Dysfunction Mol Cell Biol 2012 Dec 17 [PMID: 23249949] (IHC, WB, Mouse)
• Deschepper M, Manassero M, Oudina K et al. Proangiogenic and Prosurvival Functions of Glucose in Human Mesenchymal Stem Cells upon Transplantation. Stem Cells 2013 Mar [PMID: 23225732] (WB, Human)
• Riddell JR, Maier P, Sass SN et al. Peroxiredoxin 1 Stimulates Endothelial Cell Expression of VEGF via TLR4 Dependent Activation of HIF-1alpha PLoS One 2012 [PMID: 23185615] (IHC, WB, Human, Mouse)
• Neumeister VM, Anagnostou V, Siddiqui S et al. Quantitative Assessment of Effect of Preanalytic Cold Ischemic Time on Protein Expression in Breast Cancer Tissues J Natl Cancer Inst 2012 Oct 22 [PMID: 23090068] (IHC, Human)
• Xiao H, Li S, Zhang D et al. Separate and concurrent use of 2-deoxy-D-glucose and 3-bromopyruvate in pancreatic cancer cells. Oncol Rep. 2013 Jan [PMID: 23076497] (WB, Human)
• Shin MK, Drager LF, Yao Q et al. Metabolic Consequences of High-Fat Diet Are Attenuated by Suppression of HIF-1alpha PLoS One 2012 [PMID: 23049707] (WB, Mouse)
• Veith C, Marsh LM, Wygrecka M et al. Paxillin regulates pulmonary arterial smooth muscle cell function in pulmonary hypertension. Am J Pathol 2012 Nov [PMID: 22959909]
• Kunze R, Zhou W, Veltkamp R et al. Neuron-specific prolyl-4-hydroxylase domain 2 knockout reduces brain injury after transient cerebral ischemia. Stroke 2012 Oct [PMID: 22933585] (Mouse)
• Kottmann RM, Kulkarni AA, Smolnycki KA et al. Lactic Acid Is Elevated in Idiopathic Pulmonary Fibrosis and Induces Myofibroblast Differentiation via pH-Dependent Activation of Transforming Growth Factor-beta. Am J Respir Crit Care Med 2012 Oct 15 [PMID: 22923663]
• Hamidi T, Cano CE, Grasso D et al. Nupr1-aurora kinase A pathway provides protection against metabolic stress-mediated autophagic-associated cell death. Clin Cancer Res 2012 Oct [PMID: 22899799]
• Ji K, Tsirka SE. Inflammation modulates expression of laminin in the central nervous system following ischemic injury. J Neuroinflammation 2012 Jul [PMID: 22759265]
• Pohjoismaki JL, Boettger T, Liu Z, Goffart S, Szibor M, Braun T. Oxidative stress during mitochondrial biogenesis compromises mtDNA integrity in growing hearts and induces a global DNA repair response. Nucleic Acids Res. 2012 Apr 15. [PMID: 22508755] (WB, Rat)
• Wu MH, Lu CW, Chang FM, Tsai SJ. Estrogen receptor expression affected by hypoxia inducible factor-1 alpha in stromal cells from patients with endometriosis Taiwan J Obstet Gynecol 2012 Mar [PMID: 22482968] (WB, Human)
• Gayed BA, O'Malley KJ, Pilch J, Wang Z. Digoxin Inhibits Blood Vessel Density and HIF-1a Expression in Castration-Resistant C4-2 Xenograft Prostate Tumors. Clin Transl Sci;;5(1):39-42. doi: 10.1111/j.1752-8062.2011.00376.x. 2012 Feb. [PMID: 22376255]
• Shiau AL, Shen YT, Hsieh JL et al. Scutellaria barbata inhibits angiogenesis through downregulation of HIF-1 alpha in lung tumor. Environ Toxicol 2014 Apr [PMID: 22331677]
• Chiusa M, Hool SL, Truetsch P, Djafarzadeh S, Jakob SM, Seifriz F, Scherer SJ, Suter TM, Zuppinger C, Zbinden S. Cancer therapy modulates VEGF signaling and viability in adult rat cardiac microvascular endothelial cells and cardiomyocytes. J Mol Cell Cardiol. 2012 Feb 3. [PMID: 22326847]
• Zhang L, Das P, Schmolke M et al. Inhibition of pyrimidine synthesis reverses viral virulence factor-mediated block of mRNA nuclear export. J Cell Biol 2012 Feb [PMID: 22312003]
• Copple BL, Kaska S, Wentling C. Hypoxia-inducible Factor Activation in Myeloid Cells Contributes to the Development of Liver Fibrosis in Cholestatic Mice. The Journal of Pharmacology and Experimental Therapeutics. 2012 Jan 23. [PMID: 22271822]
• Barta P, Van Pelt C, Men T et al. Enhancement of lung tumorigenesis in a Gprc5a Knockout mouse by chronic extrinsic airway inflammation Mol Cancer 2012 Jan 12 [PMID: 22239913] (IHC, Mouse)
• Ko SH, Nauta A, Morrison SD et al. Antimycotic ciclopirox olamine in the diabetic environment promotes angiogenesis and enhances wound healing. PLoS ONE 20116(11). [PMID: 22125629]
• Hong SJ, Jin da P, Buck DW 2nd et al. Impaired response of mature adipocytes of diabetic mice to hypoxia. Exp Cell Res 2011 Oct [PMID: 21777581]
• Rabie T, Kunze R, Marti HH. Impaired hypoxic response in senescent mouse brain. Int J Dev Neurosci. 2011 Jun 16. [PMID: 21704147]
• Jantsch J, Wiese M, Schodel J et al. Toll-like receptor activation and hypoxia use distinct signaling pathways to stabilize hypoxia-inducible factor 1{alpha} (HIF1A) and result in differential HIF1A-dependent gene expression. J Leukoc Biol. 2011 Jun 17. [PMID: 21685248]
• Pascal LE, Ai J, Rigatti LH et al. EAF2 loss enhances angiogenic effects of Von Hippel-Lindau heterozygosity on the murine liver and prostate. Angiogenesis 2011 Sept [PMID: 21638067] (WB, Mouse)
• Guadall A, Orriols M, Alcudia JF et al. Hypoxia-induced ROS signaling is required for LOX up-regulation in endothelial cells Front Biosci 2011 Jun 1 [PMID: 21622104] (WB, Human)
• Jiang X, Khan MA, Tian W et al. Adenovirus-mediated HIF-1alpha gene transfer promotes repair of mouse airway allograft microvasculature and attenuates chronic rejection Clin Invest 2011 Jun [PMID: 21606594]
• Duan LJ, Takeda K, Fong GH et al. Prolyl hydroxylase domain protein 2 (PHD2) mediates oxygen-induced retinopathy in neonatal mice Am J Pathol. 2011 Apr [PMID: 21435465] (WB, Mouse)
• Dunford ECE, Herbst EAF, Jeoung NH et al. Pyruvate dehydrogenase (PDH) activation during in vitro muscle contractions in PDH kinase 2 knockout mice: the effect of PDH kinase 1 compensation. Am J Physiol Regul Integr Comp Physiol. 2011 Mar 16. [PMID: 21411764]
• Wolff NC, Vega-Rubin-de-Celis S, Xie XJ et al. Cell-type-dependent regulation of mTORC1 by REDD1 and the tumor suppressors TSC1/TSC2 and LKB1 in response to hypoxia. Mol Cell Biol 2011 May [PMID: 21383064]
• Nagao K, Oka K. HIF-2 directly activates CD82 gene expression in endothelial cells. Biochem Biophys Res Commun;407(1):260-5. 2011 Apr 1. [PMID: 21382346]
• Camacho M, Rodriguez C, Guadall A et al. Hypoxia upregulates PGI-synthase and increases PGI2 release in human vascular cells exposed to inflammatory stimuli J Lipid Res 2011 Apr [PMID: 21296955] (WB, Human)
• Li Q, Michaud M, Canosa S, Kuo A, Madri JA. GSK-3?: a signaling pathway node modulating neural stem cell and endothelial cell interactions. Angiogenesis;14(2):173-85. 2011 May. [PMID: 21253820] (WB, Mouse)
• Huang M, Wu JC. Molecular imaging of RNA interference therapy targeting PHD2 for treatment of myocardial ischemia. Methods Mol Biol;709:211-21. 2011 [PMID: 21194030] (WB, Mouse)
• Doran DM, Kulkarni-Datar K, Cool DR et al. Hypoxia Activates Constitutive Luciferase Reporter Constructs. Biochimie. 2010 Oct 21. [PMID: 20971156]
• Pirkmajer S, Filipovic D, Mars T et al. HIF-1alpha response to hypoxia is functionally separated from the glucocorticoid stress response in the in vitro regenerating human skeletal muscle. Am J Physiol Regul Integr Comp Physiol 2010 Dec [PMID: 20943857] (WB, Human)
• Xu D, Yao Y, Lu L et al. Plk3 functions as an essential component of the hypoxia regulatory pathway by direct phosphorylation of HIF-1alpha. J Biol Chem 2010 Dec [PMID: 20889502]
• Copple BL, Bai S, Burgoon LD, Moon J-O. Hypoxia-inducible factor-1[alpha] regulates the expression of genes in hypoxic hepatic stellate cells important for collagen deposition and angiogenesis. Liver Int. [PMID: 20880076]
• Welford SM, Dorie MJ, Li X et al. Renal oxygenation suppresses VHL loss-induced senescence that is caused by increased sensitivity to oxidative stress. Mol Cell Biol 2010 Oct [PMID: 20679489]
• Gimm T, Wiese M, Teschemacher B et al. Hypoxia-inducible protein 2 is a novel lipid droplet protein and a specific target gene of hypoxia-inducible factor-1 FASEB J 2010 Nov [PMID: 20624928] (WB, Human)
• Tam KK, Russell DL, Peet DJ et al. Hormonally regulated follicle differentiation and luteinization in the mouse is associated with hypoxia inducible factor activity. Mol Cell Endocrinol 2010 Oct 7 [PMID: 20600586] (WB, Mouse)
• Mikheyev AS, Vo T, Wee B et al. Rapid microsatellite isolation from a butterfly by de novo transcriptome sequencing: performance and a comparison with AFLP-derived distances. PLoS One 2010 [PMID: 20585453]
• Copple BL, Bai S, Moon JO. Hypoxia-inducible factor-dependent production of profibrotic mediators by hypoxic Kupffer cells. Hepatol Res;40(5):530-9. 2010 May. [PMID: 20412331]
• Lu ZH, Shvartsman MB, Lee AY et al. Mammalian target of rapamycin activator RHEB is frequently overexpressed in human carcinomas and is critical and sufficient for skin epithelial carcinogenesis. Cancer Res 2010 Apr [PMID: 20388784]
• Serra-Perez A, Planas AM, Nunez-O'Mara A et al. Extended ischemia prevents HIF1alpha degradation at reoxygenation by impairing prolyl-hydroxylation: role of Krebs cycle metabolites. J Biol Chem 2010 Jun 11 [PMID: 20368331]
• Costa B, Dettori D, Lorenzato A et al. Fumarase tumor suppressor gene and MET oncogene cooperate in upholding transformation and tumorigenesis. FASEB J 2010 Aug [PMID: 20354140]
• Kuphal S, Winklmeier A, Warnecke C, Bosserhoff AK. Constitutive HIF-1 activity in malignant melanoma Eur J Cancer 2010 Apr [PMID: 20185296] (WB, Human)
• Clever JL, Sakai Y, Wang RA et al. Inefficient skeletal muscle repair in inhibitor of differentiation knockout mice suggests a crucial role for BMP signaling during adult muscle regeneration. Am J Physiol Cell Physiol;298(5):C1087-99. 2010 May. [PMID: 20181926] (WB, Mouse)
• Copple BL. Hypoxia stimulates hepatocyte epithelial to mesenchymal transition by hypoxia-inducible factor and transforming growth factor-beta-dependent mechanisms Liver Int 2010 May [PMID: 20158611] (WB, Mouse)
• el Filali M, Missotten GS, Maat W et al. Regulation of VEGF-A in uveal melanoma. Invest Ophthalmol Vis Sci 2010 May [PMID: 20042655]
• Porcelli AM, Ghelli A, Ceccarelli C et al. The genetic and metabolic signature of oncocytic transformation implicates HIF1alpha destabilization. Hum Mol Genet 2010 Mar [PMID: 20028790]
• Samarin J, Wessel J, Cicha I et al. FoxO proteins mediate hypoxic induction of connective tissue growth factor in endothelial cells. J Biol Chem 2010 Feb 12 [PMID: 20018872]
• S K Martin, P Diamond, S A Williams et al. Hypoxia-inducible factor-2 is a novel regulator of aberrant CXCL12 expression in multiple myeloma plasma cells. Haematologica; 95 (5): 776-784. 2010 [PMID: 20015878]
• Volke M, Gale DP, Maegdefrau U et al. Evidence for a lack of a direct transcriptional suppression of the iron regulatory peptide hepcidin by hypoxia-inducible factors. PLoS One 2009 Nov 18 [PMID: 19924283] (WB, Human)
• Tanaka T, Wiesener M, Bernhardt W et al. The human HIF (hypoxia-inducible factor)-3alpha gene is a HIF-1 target gene and may modulate hypoxic gene induction. Biochem J;424(1):143-51. 2009 Oct 23. [PMID: 19694616]
• Loinard C, Ginouves A, Vilar J et al. Inhibition of prolyl hydroxylase domain proteins promotes therapeutic revascularization. Circulation 2009 Jul [PMID: 19546390]
• Gao Peng, Zhu Yiqian, Ling Feng et al. Nonischemic cerebral venous hypertension promotes a pro-angiogenic stage through HIF-1 downstream genes and leukocyte-derived MMP-9. J Cereb Blood Flow Metab 2009 [PMID: 19471278] (IHC, Mouse)
• Huang C, Hales BF. Teratogen responsive signaling pathways in organogenesis stage mouse limbs. Reprod Toxicol 2009 Apr [PMID: 19429390] (WB, Primate, Mouse)
• Copple BL, Bustamante JJ, Welch TP et al. Hypoxia-inducible factor-dependent production of profibrotic mediators by hypoxic hepatocytes. Liver Int 2009 Aug [PMID: 19302442] (WB, ICC/IF, Mouse)
• Xiao W, Ai J, Habermacher G et al. U19/Eaf2 Binds to Stabilizes von Hippel-Lindau Protein. Cancer Res;69(6):2599-2606. 2009 [PMID: 19258512] (WB)
• Houten SM, Chegary M, Te Brinke H et al. Pyruvate dehydrogenase kinase 4 expression is synergistically induced by AMP-activated protein kinase and fatty acids. Cell Mol Life Sci 2009 Apr [PMID: 19224132] (WB, Rat)
• Lomb DJ, Desouza LA, Franklin JL et al. Prolyl hydroxylase inhibitors depend on extracellular glucose HIF-2{alpha} to inhibit cell death caused by NGF deprivation: evidence that HIF-2{alpha} has a role in NGF-promoted survival of sympathetic neurons. Mol Pharmacol;75(5):1198-209. 2009 May. [PMID: 19204094] (WB)
• Vasseur S, Afzal S, Tardivel-Lacombe J et al. DJ-1/PARK7 is an important mediator of hypoxia-induced cellular responses. Proc Natl Acad Sci U S A 2009 Jan [PMID: 19144925]
• Moon J-OK, Welch TP, Gonzalez FJ et al. Reduced liver fibrosis in hypoxia-inducible factor-1{alpha}-deficient mice. Am J Physiol Gastrointest Liver Physiol;296(3):G582-592. 2009 [PMID: 19136383] (WB, ICC/IF, IHC-Fr, Mouse)
• Lu CW, Lin SC, Chen KF et al. Induction of pyruvate dehydrogenase kinase-3 by hypoxia-inducible factor-1 promotes metabolic switch and drug resistance. J Biol Chem;283(42):28106-14. 2008 Oct 17. [PMID: 18718909] (WB, Human)
• Shui YB, Arbeit JM, Johnson RS et al. HIF-1: an age-dependent regulator of lens cell proliferation. Invest Ophthalmol Vis Sci 2008 Nov [PMID: 18586877]
• Yang Y, Bai J, Shen R et al. Polo-like kinase 3 functions as a tumor suppressor and is a negative regulator of hypoxia-inducible factor-1 alpha under hypoxic conditions. Cancer Res 2008 Jun [PMID: 18519666]
• Wu S, Nishiyama N, Kano MR et al. Enhancement of angiogenesis through stabilization of hypoxia-inducible factor-1 by silencing prolyl hydroxylase domain-2 gene. Mol Ther 2008 Jul [PMID: 18500250]
• Kanasaki K, Palmsten K, Sugimoto H et al. Deficiency in catechol-O-methyltransferase and 2-methoxyoestradiol is associated with pre-eclampsia. Nature 2008 Jun 19 [PMID: 18469803]
• Warnecke C, Weidemann A, Volke M et al. The specific contribution of hypoxia-inducible factor-2alpha to hypoxic gene expression in vitro is limited and modulated by cell type-specific and exogenous factors. Exp Cell Res 2008 Jun 10 [PMID: 18420194] (Human)
• Vangeison G, Carr D, Federoff HJ et al. The good, the bad, and the cell type-specific roles of hypoxia inducible factor-1 alpha in neurons and astrocytes. J Neurosci;28(8):1988-93. 2008 Feb 20. [PMID: 18287515] (WB, Mouse)
• Sun L, Marti HH, Veltkamp R. Hyperbaric oxygen reduces tissue hypoxia and hypoxia-inducible factor-1 alpha expression in focal cerebral ischemia. Stroke;39(3):1000-6. 2008 Mar. [PMID: 18239183] (ICC/IF, IHC-P, Mouse)
• Becker CM, Rohwer N, Funakoshi T et al. 2-methoxyestradiol inhibits hypoxia-inducible factor-1{alpha} and suppresses growth of lesions in a mouse model of endometriosis. Am J Pathol;172(2):534-44. 2008 Feb. [PMID: 18202195] (WB, IHC-P, Mouse)
• Scortegagna M, Cataisson C, Martin RJ et al. HIF-1alpha regulates epithelial inflammation by cell autonomous NFkappaB activation and paracrine stromal remodeling. Blood 2008 Apr [PMID: 18199827]
• May D, Gilon D, Djonov V et al. Transgenic system for conditional induction and rescue of chronic myocardial hibernation provides insights into genomic programs of hibernation. Proc Natl Acad Sci U S A;105(1):282-7. 2008 Jan 8. [PMID: 18162550] (IHC-P, Mouse)
• Takeda K, Aguila HL, Parikh NS et al. Regulation of adult erythropoiesis by prolyl hydroxylase domain proteins. Blood;111(6):3229-35. 2008 Mar 15. [PMID: 18056838] (WB, Mouse)
• Friese MA, Craner MJ, Etzensperger R et al. Acid-sensing ion channel-1 contributes to axonal degeneration in autoimmune inflammation of the central nervous system. Nat Med 2007 Dec [PMID: 17994101] (Mouse)
• Glassford AJ, Yue P, Sheikh AY et al. HIF-1 regulates hypoxia- and insulin-induced expression of apelin in adipocytes. Am J Physiol Endocrinol Metab;293(6):E1590-6. 2007 Dec. [PMID: 17878221] (WB, Mouse)
• Palmer LA, Doctor A, Chhabra P et al. S-nitrosothiols signal hypoxia-mimetic vascular pathology. J Clin Invest;117(9):2592-601. 2007 Sep. [PMID: 17786245] (WB, Mouse)
• Takeda K, Cowan A, Fong GH et al. Essential role for prolyl hydroxylase domain protein 2 in oxygen homeostasis of the adult vascular system Circulation. 2007 Aug 14 [PMID: 17646578] (WB, Mouse)
• Chen HL, Pistollato F, Hoeppner DJ et al. Oxygen tension regulates survival and fate of mouse central nervous system precursors at multiple levels. Stem Cells;25(9):2291-301. 2007 Sep. [PMID: 17556599] (ELISA, Mouse)
• van der Bilt JD, Soeters ME, Duyverman AM et al. Perinecrotic hypoxia contributes to ischemia/reperfusion-accelerated outgrowth of colorectal micrometastases. Am J Pathol 2007 Apr [PMID: 17392176] (Mouse)
• Nagano M, Yamashita T, Hamada H et al. Identification of functional endothelial progenitor cells suitable for the treatment of ischemic tissue using human umbilical cord blood. Blood;110(1):151-60. 2007 Jul 1. [PMID: 17379743] (WB, Human)
• van de Sluis B, Muller P, Duran K et al. Increased activity of hypoxia-inducible factor 1 is associated with early embryonic lethality in Commd1 null mice. Mol Cell Biol 2007 Jun [PMID: 17371845]
• Bedogni B, Welford SM, Kwan AC et al. Inhibition of phosphatidylinositol-3-kinase and mitogen-activated protein kinase kinase 1/2 prevents melanoma development and promotes melanoma regression in the transgenic TPRas mouse model. Mol Cancer Ther 2006 Dec [PMID: 17172409]
• Welford SM, Bedogni B, Gradin K et al. HIF1alpha delays premature senescence through the activation of MIF. Genes Dev 2006 Dec [PMID: 17142669]
• Kim WY, Safran M, Buckley MR et al. Failure to prolyl hydroxylate hypoxia-inducible factor alpha phenocopies VHL inactivation in vivo. EMBO J 2006 Oct 4 [PMID: 16977322]
• Takeda K, Ho VC, Takeda H 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)
• Litz J, Krystal GW. Imatinib inhibits c-Kit-induced hypoxia-inducible factor-1alpha activity and vascular endothelial growth factor expression in small cell lung cancer cells. Mol Cancer Ther 2006 Jun [PMID: 16818499] (Human)
• Martin-Oliva D, Aguilar-Quesada R, O'valle F et al. Inhibition of poly(ADP-ribose) polymerase modulates tumor-related gene expression, including hypoxia-inducible factor-1 activation, during skin carcinogenesis. Cancer Res 2006 Jun [PMID: 16740713]
• Hu CJ, Iyer S, Sataur A 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 2006 May [PMID: 16611993] (WB)
• Zampino M, Yuzhakova M, Hansen J et al. Sex-related dimorphic response of HIF-1 alpha expression in myocardial ischemia. Am J Physiol Heart Circ Physiol 2006 Aug [PMID: 16603692]
• Hsu T, Adereth Y, Kose N et al. Endocytic function of von Hippel-Lindau tumor suppressor protein regulates surface localization of fibroblast growth factor receptor 1 and cell motility. J Biol Chem;281(17):12069-80. 2006 Apr 28. [PMID: 16505488] (WB, Human)
• Litz J, Krystal GW. Imatinib inhibits c-Kit-induced hypoxia-inducible factor-1alpha activity and vascular endothelial growth factor expression in small cell lung cancer cells. Mol Cancer Ther;5(6):1415-22. 2006 Jun. [PMID: 1681849] (WB, Human)

Reviews for HIF-1 alpha Antibody (NB100-449) (28)

Average Rating: 4.3

(Based on 28 reviews)

We have 28 reviews tested in 5 species: Human, Mouse, Rat, Goat, Other.

reviewed by: Anonymous WB Mouse 05/08/2020

Summary

• Application: Western Blot
• Sample Tested: 3T3-L1 mouse embryonic fibroblast adipose-like cell line
• Species: Mouse
• Lot: lot A3

Comments

• Comments: skimmed milk 5%
  primary : 1:2000
  secondary: 1: 200000

reviewed by: Anonymous IHC-P Mouse 06/07/2019

Summary

• Application: Immunohistochemistry-Paraffin
• Sample Tested: PDX mouse
• Species: Mouse
• Lot: Lot A3

Comments

• Comments: Working Dilution 1:100, EDTA pH 9.0; detection kit Polymer-HRP.

reviewed by: an shiyu WB Goat 05/15/2018

Summary

• Application: Western Blot
• Sample Tested: laydig cell
• Species: Goat
• Lot: NBP1-04676SS

Comments

• Comments: dilution:1:500

reviewed by: Anonymous WB Mouse 07/17/2017

Summary

• Application: Western Blot
• Sample Tested: BMDM
• Species: Mouse
• Lot: A5

reviewed by: Anonymous IP Rat 04/24/2017

Summary

• Application: Immunoprecipitation
• Sample Tested: PC-12 rat adrenal pheochromocytoma cell line
• Species: Rat
• Lot: A6

reviewed by: Anonymous IF Mouse 01/12/2016

Summary

• Application: Immunofluorescence
• Sample Tested: primary bone marrow derived macrophages
• Species: Mouse

Blocking

• Blocking Details: -

Details

• Detection Notes: -

Comments

• Comments: Murine primary bone marrow derived macrophages probed with rabbit HIF1a antibody (1:100) in blocking buffer at 4°C overnight, secondary goat anti-rabbit AF647 (1:250) for 2 h at ambient temperature, Cells were mounted with Vectashield containing DAPI for imaging. Image (top right) derived from merge of DAPI and AF647 staining. White arrows point at cells containing nuclear HIF1a. Unstained and secondary antibody controls shown in bottom panels, Magnification, 63X

reviewed by: Anonymous WB Human 09/07/2015

Summary

• Application: Western Blot
• Sample Tested: Human cancer cell whole cell lysate
• Species: Human

Blocking

• Blocking Details: Blocking with 5% non-fat milk for 30 min

Primary Anitbody

• Dilution Ratio: 1:1000, overnight

Secondary Antibody

• Secondary Description: Donkey anti-rabbit IgG, HRP
• Secondary Concentration: 1:2000

Details

• Detection Notes: Regular Protocol

reviewed by: Zachary Patinkin Simple Western Human 06/09/2015

Summary

• Application: Simple Western
• Sample Tested: Human mesenchymal stem cell whole cell lysate
• Species: Human
• Lot: A5

Blocking

• Blocking Details: Protein Simple Antibody Diluent, 5 min, 25C

Primary Anitbody

• Dilution Ratio: 1:200, 30 minutes, Protein Simple Antibody Diluent

Secondary Antibody

• Secondary Description: Protein Simple Anti-Rabbit Secondary
• Secondary Manufacturer Cat#: 042-206
• Secondary Concentration: undiluted

Details

• Detection Notes: WES multi-image analysis

reviewed by: Enrica Borsi WB Human 01/09/2015

Summary

• Application: Western Blot
• Sample Tested: Myeloma cells whole cell lysate
• Species: Human

Blocking

• Blocking Details: Blocking buffer Milk 5%

Details

• Detection Notes: Western blot

reviewed by: Anonymous WB Human 12/12/2014

Summary

• Application: Western Blot
• Sample Tested: See PMID 22923663
• Species: Human

Blocking

• Blocking Details: See PMID 22923663

Details

• Detection Notes: See PMID 22923663

Comments

• Comments: Published in PMID: 22923663

reviewed by: Ming Lu WB Human 08/15/2014

Summary

• Application: Western Blot
• Sample Tested: Human pancreatic cancer cell lysates
• Species: Human
• Lot: A4
• Comments: Although I got my result as expected, this antibody has weak bands and dirty background in my western blotting.

Blocking

• Blocking Details: 2% cold fish geletin in TBS buffer, room temperature for 1hour

Primary Anitbody

• Dilution Ratio: 1:1000 overnight at 4C in blocking buffer

Secondary Antibody

• Secondary Description: Goat anti-rabbit IgG, HRP
• Secondary Manufacturer Cat#: Cell Signaling, #7074
• Secondary Concentration: 1:5000

Details

• Detection Notes: 4 min exposure shows band at correct MW.

Comments

• Comments: Although I got my result as expected, this antibody has weak bands and dirty background in my western blotting.

reviewed by: Xiuquan Luo WB Human 07/18/2014

Summary

• Application: Western Blot
• Sample Tested: Caki-1 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

Details

• Detection Notes: ECL, exposured for 1 min

reviewed by: Anonymous WB Mouse 07/05/2014

Summary

• Application: Western Blot
• Sample Tested: large intestinal lamina propria leukocytes (LPLs).
• Species: Mouse

reviewed by: Paramananda Saikia WB Mouse 06/30/2014

Summary

• Application: Western Blot
• Sample Tested: Mouse
• Species: Mouse
• Lot: A4

Blocking

• Blocking Details: 5% bsa

Primary Anitbody

• Dilution Ratio: 1:1500

Secondary Antibody

• Secondary Description: goat anti-rabbit

reviewed by: Alshaimaa Adawy WB Human 04/29/2014

Summary

• Application: Western Blot
• Sample Tested: nuclear fractions from T47-D beast cancer cell line
• Species: Human
• Lot: A5

Blocking

• Blocking Details: blocking: 5% BSA for 1 hour at room temprature

Primary Anitbody

• Dilution Ratio: 1:1000, overnight at 4 C in BSA

Secondary Antibody

• Secondary Description: Anti-rabbit IgG, HRP-linked Antibody
• Secondary Manufacturer Cat#: 7074s, cell signaling
• Secondary Concentration: 1:5000

Details

• Detection Notes: Vilber Lourmat imager, using home made ECL, exposure time 3 minutes

reviewed by: Anonymous WB Other 03/14/2014

Summary

• Application: Western Blot
• Species: Other
• Lot: A4
• Pub Med ID: 21193390
• File: View PDF

reviewed by: Lidan Zhao WB Other 08/26/2013

Summary

• Application: Western Blot
• Sample Tested: pig satellite cell nuclear protein
• Species: Other
• Comments: I first ordered NB100-105 and tried several times, but it didn't work with my samples. I contacted technical support and they sent me a free vial of NB100-449 as a replacement. I found that this antibody worked well both on the positive control and my pig samples.

Blocking

• Blocking Details: 5% milk in TBST

Primary Anitbody

• Dilution Ratio: 1:500 in 5% milk, 4 C overnight

Secondary Antibody

• Secondary Description: IRDye 800CW Goat anti-Rabbit IgG
• Secondary Concentration: 1:20000

Details

• Detection Notes: IRDye conjugated secondary scanned by Licor Odyssey system. Used Novus COS-7 nuclear extracts (NB800-PC26) as positive control

Comments

• Comments: I first ordered NB100-105 and tried several times, but it didn't work with my samples. I contacted technical support and they sent me a free vial of NB100-449 as a replacement. I found that this antibody worked well both on the positive control and my pig samples.

reviewed by: Anonymous WB Human 11/05/2012

Summary

• Application: Western Blot
• Sample Tested: human cell lysates
• Species: Human

Blocking

• Blocking Details: 5% Non fat milk

Primary Anitbody

• Dilution Ratio: 1:500

Secondary Antibody

• Secondary Description: anti-rabbit-HRP
• Secondary Manufacturer Cat#: Sigma
• Secondary Concentration: 1:1000

Details

• Detection Notes: ECL

reviewed by: Annie Curtis WB Mouse 08/24/2012

Summary

• Application: Western Blot
• Sample Tested: mouse peritoneal and bone marrow derived macrophages
• Species: Mouse
• Lot: A4
• Comments: We tried numerous HIF1alpha antibodies with little success, we finally got this one to work but one thing that is necessary is that you need to lyse your cells directly in loading buffer. We used about 100uL of Laemmli buffer per 6 well dish, then boil immediately after lysing. Samples can then be either loaded directly onto the gel or stored at -20. However, we did find that if samples were stored for too long at -20 then we did lose the ability to detect the protein.

Blocking

• Blocking Details: blocked in 5% milk for 1 hour at room temperature

Primary Anitbody

• Dilution Ratio: 1:1000 overnight at 4deg C, it has to be an overnight incubation to work

Secondary Antibody

• Secondary Description: Goat anti-rabbit IgG, HRP
• Secondary Concentration: 1: 1000

Details

• Detection Notes: ECL exposure of 2 minutes needed

Comments

• Comments: We tried numerous HIF1alpha antibodies with little success, we finally got this one to work but one thing that is necessary is that you need to lyse your cells directly in loading buffer. We used about 100uL of Laemmli buffer per 6 well dish, then boil immediately after lysing. Samples can then be either loaded directly onto the gel or stored at -20. However, we did find that if samples were stored for too long at -20 then we did lose the ability to detect the protein.

reviewed by: kimberley tam WB 08/07/2012

Summary

• Application: Western Blot
• Lot: A2
• Special Applications: Excellent antibody. Tested with mouse
• Pub Med ID: 20600586

reviewed by: Anonymous WB 05/09/2012

Summary

• Application: Western Blot
• Pub Med ID: 16611993
• File: View PDF

reviewed by: Anonymous WB Rat 01/03/2012

Summary

• Application: Western Blot
• Sample Tested: PC12 cell whole lysate, Sample Amount: 25ug
• Species: Rat
• Lot: A4
• Comments: This antibody could detect HIF-1a but weak.

Blocking

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

Primary Anitbody

• Dilution Ratio: Dilution Ratio: 1:1000, Incubation Dilution Buffer: blocking buffer, Incubation Time: overnight, Incubation Temp: 4 C

Secondary Antibody

• Secondary Description: Secondary Ab: anti-Rabbit IgG, Secondary Ab Dilution Ratio: 1:5000

Details

• Detection Notes: Detection Method: ECL Plus

Comments

• Comments: This antibody could detect HIF-1a but weak.

reviewed by: Anonymous WB Human 06/08/2011

Summary

• Application: Western Blot
• Sample Tested: HUVEC whole cell lysate, Sample Amount: 20 microg
• Species: Human
• Comments: Western blot analysis showing that the increase in HIF-1alpha levels triggered by hypoxia in HUVEC is prevented by a specific siRNA against HIF-1alpha (siHIFa).(N: Normoxia, H: Hypoxia; siRAND: Random siRNA
• Pub Med ID: 21622104
• File: View PDF

Blocking

• Blocking Details: Blocking Buffer: 5%Milk in TBST, Blocking Time: overnight, Blocking Temp: 4 degrees Celsius

Primary Anitbody

• Dilution Ratio: Dilution Ratio: 1:500, Incubation Dilution Buffer: 5% Milk in TBST, Incubation Time: 1 hour, Incubation Temp: room temperature

Secondary Antibody

• Secondary Description: Secondary Ab: anti-rabbit DAKO, Secondary Ab Dilution Ratio: 1:2000
• Secondary Manufacturer Cat#: P0448

Details

• Detection Notes: Method: SuperSignal West, Time: 3min, Controls: Hypoxia/normoxia in random transfected cells, Bands: Multiband at 110kDa

Comments

• Comments: Western blot analysis showing that the increase in HIF-1alpha levels triggered by hypoxia in HUVEC is prevented by a specific siRNA against HIF-1alpha (siHIFa).(N: Normoxia, H: Hypoxia; siRAND: Random siRNA

reviewed by: Anonymous WB Mouse 12/22/2010

Summary

• Application: Western Blot
• Sample Tested: cerebellar neurons
• Species: Mouse
• Comments: This Hif1 alpha antibody is the only one we have found to work consistently well. We use it 1:500 in 1% BSA and reuse it for 4-5 times. We get a a doublet, the expected band at around 100KDa and another band at about 130KDa or so (right below the 150KDa marker.)

Blocking

• Blocking Details: Blocking Buffer: 3% BSA in TBST, Blocking Time: 1 hour, Blocking Temp: Room temperature

Primary Anitbody

• Dilution Ratio: Primary Ab Dilution Ratio: 1:500, Primary Ab Incubation Time: overnight, Primary Ab Incubation Temp: 4 degrees Celsius

Secondary Antibody

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

Details

• Detection Notes: Detection Method: ECL, Exposure Time: 5 minutes

Comments

• Comments: This Hif1 alpha antibody is the only one we have found to work consistently well. We use it 1:500 in 1% BSA and reuse it for 4-5 times. We get a a doublet, the expected band at around 100KDa and another band at about 130KDa or so (right below the 150KDa marker.)

reviewed by: William Lin IP Human 12/03/2010

Summary

• Application: Immunoprecipitation
• Sample Tested: HEK 293
• Species: Human
• Lot: A3
• Comments: I used BD mouse anti-human HIF1 antibody for WB because I didn't want to detect the rabbit IgG from the NB100-449 antibody during the IP. Lysate Amount: 1mg, Lysate Preclear: agarose A, Diluted Lysate Concentration: 2ug/ul, Ab Dilution Ratio: 1:100, Protein Capture: agarose A, Collection Method: 4000 rpm centrifugation, Protein Reduction: 2X sample buffer, with SDS/bME, Bead Removal: 4min, 4C

Blocking

• Blocking Details: Blocking Buffer: 5% milk in TBST, Blocking Time: 2 hours, Blocking Temp: 4 degrees Celsius

Primary Anitbody

• Dilution Ratio: Incubation Dilution Buffer: 5% milk/TBST, Dilution Ratio: 1:2000, Incubation Time: overnight, Incubation Temp: 4 C

Secondary Antibody

• Secondary Description: Secondary Ab: anti-human IgG-HRP, Secondary Ab Dilution: 1:10000
• Secondary Manufacturer Cat#: promega

Details

• Detection Notes: Method: ECL, Exposure Time: 2 min, Pos.Ctrl: DFO treated, Neg.Ctrl: no treatment, Specific Bands: 116kDa

Comments

• Comments: I used BD mouse anti-human HIF1 antibody for WB because I didn't want to detect the rabbit IgG from the NB100-449 antibody during the IP. Lysate Amount: 1mg, Lysate Preclear: agarose A, Diluted Lysate Concentration: 2ug/ul, Ab Dilution Ratio: 1:100, Protein Capture: agarose A, Collection Method: 4000 rpm centrifugation, Protein Reduction: 2X sample buffer, with SDS/bME, Bead Removal: 4min, 4C

reviewed by: Anonymous WB Other 03/11/2010

Summary

• Application: Western Blot
• Sample Tested: Colon Cancer Cell lines, Sample Amount: 50ug
• Species: Other
• Lot: A2
• Comments: The lanes on the left are HCT116 treated with increasing concentrations of DMOG. The lanes on the right are HCT116 Hif1-/- cells with the same treatment.

Blocking

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

Primary Anitbody

• Dilution Ratio: Dilution Ratio: 1/5000, Incubation Dilution Buffer: 5% BSA in TNS with Tween, Incubation Time: overnight, Incubation Temp: 4C

Secondary Antibody

• Secondary Description: Secondary Ab: Cell Signaling Goat Anti-Rabbit HRP, Secondary Ab Dilution Ratio: 1/10000

Details

• Detection Notes: Detection Method: ECL Plus, Specific Bands: 120 kDa

Comments

• Comments: The lanes on the left are HCT116 treated with increasing concentrations of DMOG. The lanes on the right are HCT116 Hif1-/- cells with the same treatment.

reviewed by: Anonymous WB Mouse 12/21/2009

Summary

• Application: Western Blot
• Sample Tested: human tumor model mouse, Sample Amount: 12ug
• Species: Mouse
• Comments: Nuclear protein extracts: Chen et al. J Cell Biochem 2001;82(3):512-521.

Blocking

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

Primary Anitbody

• Dilution Ratio: Dilution Ratio: 1/500, Incubation Dilution Buffer: TBST with 5% milk, Incubation Time: overnight, Incubation Temp: 4 C

Secondary Antibody

• Secondary Description: Secondary Ab: Anti-rabbit IgG HRP-linked, Cell Signaling #7047, Secondary Ab Dilution Ratio: 1/3000

Details

• Detection Notes: Detection Method: ECL, Exposure Time: 5 - 10 min., Specific Bands: 100 kDa

Comments

• Comments: Nuclear protein extracts: Chen et al. J Cell Biochem 2001;82(3):512-521.

reviewed by: Anonymous WB Mouse 04/08/2009

Summary

• Application: Western Blot
• Sample Tested: Mouse Neuroblastoma, Sample Amount: 25ug
• Species: Mouse
• Lot: A2
• Comments: "Load:
  Empty Vector Normoxia
  Empty Vector Hypoxia</p><p>Hif1a over expression Normoxia
  Hif1a over expression Hypoxia"

Blocking

• Blocking Details: Blocking Buffer: 5% NFDM with TBS and.5% Tween 20, Blocking Time: 90 minutes, Blocking Temp: 25 degrees Celsius

Primary Anitbody

• Dilution Ratio: Dilution Ratio: 1/500, Incubation Dilution Buffer: 5% NFDM w/ TBS and.5% Tween 20, Incubation Time/Temp: overnight(16hrs) at 4C

Secondary Antibody

• Secondary Description: Secondary Ab: Santa Cruz Goat anti Rabbit Secondary, Secondary Ab Dilution Ratio: 1/5000

Details

• Detection Notes: Method: Chemiluminescence onto Kodak XAR film, Exposure Time: 2-15 min, Bands: 120,60 kDa(specific), 50 kDa(non-specific)

Comments

• Comments: "Load:
  Empty Vector Normoxia
  Empty Vector Hypoxia</p><p>Hif1a over expression Normoxia
  Hif1a over expression Hypoxia"

FAQs for HIF-1 alpha Antibody (NB100-449). (Showing 1 - 10 of 14 FAQs).

1. Why is there a difference between the theoretical MW for HIF1A and the observed MW for HIF-1 alpha?
   • HIF1A, like many other proteins, has post-translational modifications. Depending on the size, amount and nature of the post-translational modifications, it can cause subtle to very large changes in molecular weight.
2. Which antibody(ies) do you recommend for the detection of HIF-1a by immunohistochemistry in the sections of paraffin-embedded mouse liver samples? I would appreciate if you can give me several choices and rank them in the order of performance. My goal is to distinguish HIF upregulation by prolyl hydroxylase inhibitor in different liver cells.
   • All of our antibodies are of high quality and are well tested/validated in species/applications we list on the datasheet. However, we suggest the following four HIF-1 alpha antibodies based upon customer reviews, as well as the number of peer reviewed publications in which these products have been cited by researchers from reputed institutes. (1) HIF-1 alpha Antibody (H1alpha67) (cat# NB100-105) (cited in at least 218 peer reviewed publications) (2) HIF-1 alpha Antibody (cat# NB100-479) (cited in at least 51 peer reviewed publications) (3) HIF-1 alpha Antibody (H1alpha67) (cat# NB100-123 ) (cited in at least 38 peer reviewed publications) (4) HIF-1 alpha Antibody (cat# NB100-449) (cited in at least 31 peer reviewed publications).
3. I would like to know, does a path exist for detection of HIF 1 in venous blood before and after revascularization of the leg? 
   • We are not entirely sure if HIF-1 alpha will be present in the leg after revascularization. It may be present, but you may want to search the literature to see if this has been looked at before. If not, then this would certainly be an experiment worth doing.
4. What is the molecular weight (kDa) of protein HIF 1 alpha in western blot?
   • The theoretical molecular weight of HIF 1-alpha is ~93kDa. However, you will likely see a band between 100-120kDa due to phosphorylation.
5. We got the Hif1a (https://www.novusbio.com/products/hif-1-alpha-antibody-h1alpha67_nb100-105 ) antibody from you guys. I used the concentration that is mentioned on your website, but I am getting a band of a completely different size (~70kDa) and not the 120 kDa mentioned.
   • HIF-1 alpha is a notoriously difficult protein to work with due to its rapid degradation. Therefore, the ~70kDa bands are most likely degradation products. It is very important to lyse the cells in hypoxic conditions. We strongly recommend lysing the cells directly into the Laemmli buffer and doing that quickly, so that the exposure to oxygen is minimized.Please go through our hypoxia related FAQs, you should find them very informative:https://www.novusbio.com/support/hypoxia-and-hif-faqsAlso, running a positive control may help confirm the band specificity in your samples. You may prepare them yourself or choose some from our catalog, for example: 1) HeLa Hypoxic / Normoxic Cell Lysate (NBP2-36452)2) HeLa Hypoxic (CoCl2) / Normoxic Cell Lysate (NBP2-36450)
6. I performed several Western Blots of HIF-1 alpha with different lysis buffers, whole lysates, and cytoplasm/nuclei extractions. I can’t seem to get a good western blot (poor signal, band much lower than expected, etc.). Can someone suggest some technical considerations/tricks I should consider using?
   • A major issue that researchers working with HIF-1 alpha is degradation due to exposure to oxygen. In western blot, this results in a weaker band and/or the appearance of multiple low molecular weight bands (40-80 kDa). We recommend preparing the lysates after collection of cells/tissues as quickly as possible (on ice), preferably in a hypoxic chamber. We also recommend including a true hypoxia mimetic control (eg: cells treated with CoCl2, DMOG… etc.). The controls help distinguish your band of interest from potential degradation/dimer bands.For more troubleshooting tips and frequently asked questions regarding hypoxia/HIFs, you can refer to our hypoxia-related FAQs: https://www.novusbio.com/support/hypoxia-and-hif-faqs
7. I am doing HIF1 westerns in HIF-overexpressing mouse liver and adipose tissue using Novus antirabbit HIF1a antibody with overnight incubation. I am getting strong bands around 90kDa. I am aware that HIF theoretical molecular weight is 93kDa, but in westerns, the HIF band is usually around 120kDa according to my internet research. Can someone let me know if I’m getting the right HIF band or just some non-specific bands? Thanks.
   • (1)    HIF-1 alpha’s theoretical molecular weight is 93kDa. The post translationally modified/ubiquitinated form of HIF-1 alpha protein (fails to undergo proteasomal degradation) shows up as a band in the 110-130 kDa range on a Western blot.(2)    The dimeric protein may appear at a position above 200 kDa on non-reducing gels.(3)    Importantly, HIFs are among the most rapidly degradable proteins; therefore, sample preparation is highly important when analyzing HIF1 alpha or HIF2 alpha. When degraded, HIF-1 alpha may show up between 40-80 kDa position on Western blot. Degradation may be avoided by preparing the samples as soon as possible after collection of cells/tissues in hypoxic chamber. Notably, the tissues/cells should be kept on ice during lysate preparation and the lysates should be analyzed as soon as possible.(4)    For troubleshooting suggestions/feedback on more than 25 similar frequently asked questions, I would recommend visiting Novus page: FAQs - Hypoxia and HIFs https://www.novusbio.com/support/hypoxia-and-hif-faqs(5)    Last but not the least, Novus technical support team may be contacted at: technical@novusbio.com 
8. I have Hif1a nuclear protein extract at -80C. I am wondering if anyone knows how long it would be good for at that temperature since HIf1a is known to be degraded easily.Thank you!
   • You could try a few things to further inhibit the degradation.1) Use the protease inhibitors (if you are not already using them).2) Lyse cells into a buffer that contains SDS or LDS (eg: Laemmli's buffer), since SDS and LDS denature and inhibit proteases. Lysis may even be performed with reducing agents in the buffer (eg. DTT), but this will make your lysates unsuitable for BCA assay.3) Lysing samples rapidly ensures that the samples are instantly homogenized (it also shears DNA released by the SDS).5) Flash-freezing samples in liquid nitrogen rather than freezing at -80*C reduces the window of time for protease activity.6) Freeze samples in individual aliquots, instead of thawing the same vial multiple times.
9. I am curious to know the biochemical reactions of CoCl2 that mimic hypoxia. Is it that CoCl2 can bind any ubiquitin enzyme which regulates their degradation?
   • CoCl2 inhibits PHD enzymes (the body’s “oxygen sensors”) by replacing the Fe ion with Co, preventing these enzymes from marking HIF-1 alpha for degradation. CoCl2-based hypoxia mimetic samples are often used as positive control in HIF analysis. For more troubleshooting tips and frequently asked questions regarding hypoxia/HIFs, you can refer to our hypoxia-related FAQs: http://www.novusbio.com/support/hypoxia-and-hif-faqs
10. I am curious to know the biochemical reactions of CoCl2 that mimic hypoxia. Is it that CoCl2 can bind any ubiquitin enzyme which regulates their degradation?
    • CoCl2 inhibits PHD enzymes (the body’s “oxygen sensors”) by replacing the Fe ion with Co, preventing these enzymes from marking HIF-1 alpha for degradation. CoCl2-based hypoxia mimetic samples are often used as positive control in HIF analysis. For more troubleshooting tips and frequently asked questions regarding hypoxia/HIFs, you can refer to our hypoxia-related FAQs: https://www.novusbio.com/support/hypoxia-and-hif-faqs
11. Is cross-reactivity with HIF-2 alpha tested/predicted?
    • Although we don’t have cross-reactivity data with regards to HIF-2 alpha, we predict minimal cross-reactivity based on low sequence similarity observed from BLAST analysis between HIF-1 alpha and HIF-2 alpha.
12. Do you have any specific protocol or relative literature for HIF1a FACS staining?
    • This particular antibody has not been tested in house, so unfortunately we do not have a specific protocol for this. We have a collaborator who tested this antibody in FACS and that image is on our datasheet. With the image is a brief of their protocol: 'Flow Cytometry: HIF-1 alpha Antibody [NB100-449] - Hela cells were treated for 15 hrs with 200uM CoCl2, fixed in PFA, and permeabilized in 90% MeOH. 1 X 10^6 cells were stained with 0.125ug anti- HIF-alpha and secondary FITC-conjugated goat anti-rabbit (in a 150ul reaction). Black- treated, anti-KLH control IgG; Red- untreated, anti-HIF1-alpha; Blue- treated, anti-HIF1-alpha.'
13. I detect the 60 kDa band for HIF-1 alpha antibody (NB100-449), but no band at the right molecular weight. Do you know what this non-specific band is?
    • Most likely the 60kDa band is a degradation product of HIF1A in your samples. Since HIF-1 alpha degrades so rapidly in the presence of oxygen, our lab often sees bands at lower molecular weights. We have also experienced some non-specific banding around that molecular weight too in some tests.
14. 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.
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Bioinformatics

• Gene Symbol: HIF1A
• Entrez:
  • 3091(Human)
  • 15251(Mouse)
• Uniprot:
  • Q16665()