Technical Data

H9807-01
Clone Type
Monoclonal
Host
Mouse
Source
Human
Isotype
IgG2b
Clone Number
3C142
Grade
Affinity Purified
Applications
ChIP E FC IC IF IHC IP WB KO
Crossreactivity
Bo Hm Hu Mk Mo Po Rb Rt Sh
Shipping Temp
Blue Ice
Storage Temp
-20°C
Mouse Anti-Hypoxia Inducible Factor 1a (HIF-1a) [KO Validated]

Hypoxia contributes significantly to the pathophysiology of major categories of human disease, including myocardial and cerebral ischemia, cancer, pulmonary hypertension, congenital heart disease and chronic obstructive pulmonary disease. HIF-1 is a nuclear protein involved in mammalian oxygen homeostasis. This occurs as a posttranslational modification by prolyl hydroxylation. HIF-1 is a heterodimer composed of HIF-1 alpha and HIF-1 beta subunits. Both subunits are constantly translated. However, under normoxic conditions, human HIF-1 alpha is hydroxylated at Pro402 or Pro564 by a set of HIF prolyl hydroxylases, is polyubiquinated, and eventually degraded in proteosomes. Under hypoxic conditions, the lack of hydroxylation prevents HIF degradation and increases transcriptional activity. Therefore, the concentration of HIF-1 alpha increases in the cell. In contrast, HIF-1 beta remains stable under either condition. HIF hydroxylases provide insight into hypoxic cell responses, which may be used to help isolate therapeutic targets.

Applications
Suitable for use in Immunofluorescence/Immunocytochemistry, ELISA, Western Blot, Flow Cytometry Proximity Ligation Assay, Tissue Culture, Substratum, Ligand Activation, Knockdown Validated, Knockout Validated, Gel Super Shift Assays and Chromatin Immunoprecipitation. Other applications not tested.
Positive Control
Western Blot: 1-500 Chromatin Immunoprecipitation: 1-5ug/IP Flow Cytometry: 1:10-1:1000 ELISA: 1:100-1:2000 Immunofluorescence (IC): 1:50 Immunoprecipitation: 1:10-1:500 Immunohistochemistry (Paraffin and frozen): 1:20-1:50 Gel Super Shift Assays: 1:1-1:100 Optimal dilutions to be determined by the researcher. Nuclear extracts
Storage and Stability
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Immunogen
Fusion protein containing aa432-528 of human HIF-1 alpha. Cellular Localization: Nuclear.
Form
Supplied as a liquid in PBS, pH 7.2, 0.05% sodium azide.
Purity
Purified by Protein G affinity chromatography.
Specificity
Recognizes human HIF-1 alpha. Recognizes bands at 120kD representing HIF-1 alpha in induced tissues and cells. Multiple bands may be seen at 100-120kD representing post-translational modification of HIF-1 alpha. Species Crossreactivity: monkey, sheep, mouse, rat, ferret, porcine, hamster, bovine and rabbit.

Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.

References
1. Feldser, D., et al. Reciprocal Positive Regulation of Hypoxia-inducible Factor 1alpha and Insulin-like Growth Factor Cancer Research. 59: 3915-3918, 1999. |2. Nguyen, S.V., and Claycomb, W.C. Hypoxia Regulates the Expression of the Adrenomedullin and HIF-1 Genes in Cultured HL-1 Cardiomyocytes. Biochem. Biophys. Res. Comm. 265: 382-386, 1999. |3. Ravi, R., et al.Regulation of tumor angiogenesis by p53-induced degradation of hypoxia-inducible factor 1alpha. |Genes and Development. 14: 34-44, 2000. |4. Zhong, H. et al. Modulation of Hypoxia-inducible Factor 1alpha Expression by the Epidermal Growth Factor/Phosphatidylinositol 3-Kinase/PTEN/AKT/FRAP Pathway in Human Prostate Cancer Cells: Implications for Tumor Angiogenesis and Therapeutics. Cancer Research. 60: 1541-1545, 2000. |5. Rivard, A., et al. Age-dependent Defect in VEGF Expression is Associated with Reduced HIF-1 Activity. Journal of Biological Chemistry. July 5, 2000 (epublished). |6. Chandel, NS, et al. Reactive oxygen species generated at mitochondrial Complex III stabilize HIF-1-alpha during hypoxia: A mechanism of O2 sensing. J Biol Chem. 2000 May 31 (epublished). |7. Agani, F.H., et al. "The Role of Mitochondria in the Regulation of Hypoxia-inducible Factor 1 Expression During Hypoxia". JBC Papers in Press. Published on August 28, 2000 as Manuscript M005643200. |8. Birner, P., et al. Overexpression of Hypoxia-inducible Factor 1 alpha is a Marker for an Unfavorable Prognosis in early-Stage Invasive Cervical Cancer. Cancer Research. 60: 4693-4696, 2000. |9. Rajakumar, A., and Conrad, K. Expression, Ontogeny, and Regulation of Hypoxia-Inducible Transcription Factors in the Human Placenta. Biology of Reproduction. 63: 559-569, 2000. |10. Gardner, L.B., et al. Hypoxia inhibits G1/S transition through regulation of p27 expression. J. Biol. Chem. epublished ahead of print. Manuscript M010189200. |11. Palmer, L.A., et al. Normoxic Stabilization of Hypoxia-Inducible Factor-1 Expression and Activity: Redox-Dependent Effect of Nitrogen Oxides. Molecular Pharmacology. 58(6): 1197-1208, 2000. |12. Yu, Fang, et al. "Dynamic, Site-specific Interaction of Hypoxia-inducible Factor-1alpha with the von Hippel-Lindau Tumor Suppressor Protein" Cancer Research. 61: 4136-4142, 2001. |13. Chen, C., et al Regulation of glut1 mRNA by Hypoxia-inducible Factor-1. Journal of Biological Chemistry. 276(12): 9519-9525, 2001. |14. Birner, P., et al. Expression of Hypoxia-inducible Factor 1alpha in Epithelial Ovarian Tumors: Its Impact pn Prognosis and on response to Chemotherapy. Clinical Cancer Research. 7: 1661-1668, 2001. |15. Birner, P., et al. Expression of Hypoxia-inducible Factor 1alpha in oligodendrogliomas. Cancer. 92(1): 165-71, 2001. |16. Volm, M and R. Koomagi. Hypoxia-inducible Factor (HIF-1)and its Relationship to Apoptosis and Proliferation in Lung Cancer. |AntiCancer Research. 20: 1527-1534, 2000. |17. Schindl, M., et al. DNA repair-redox enzyme apurinic endonuclease in cervical cancer: Evaluation of redox control of HIF-1alpha and prognostic significance. |Internation Journal Oncology. 19(4): 799-802, 2001. |18. Duyndam, M., et al. Induction of Vascular Endothelial Growth Factor Expression and Hypoxia-inducible Factor 1alpha Protein by the Oxidative Stressor Arsenite. |JBC Papers in Press. Published on October 18, 2001 as Manuscript M106282200 |19. Ivan, M., et al. HIF-alpha Targeted for VHL-Mediated Destruction by Proline Hydroxylation: Implications for O2 Sensing. Science. 292(5516): 464-8, 2001. |20. Michenko, A., et al. Hypoxia-inducible factor-1 (HIF-1) mediated expression of the 6-phosphofructo-2-kinase/fructose-2,6-biophosphatase-3 (PFKBF3) gene: its possible role in the Warburg effect. JBC Papers in Press. Published on December 14, 2001 as Manuscript M110978200. |21. Makino, Y., et al. Inhibitory PAS domain protein is a negative regulator of hypoxia-inducible gene expression. |Nature. 414: 550-554, 2001. |22. Tendler, D., et al. Intersction of Interferon and Hypoxia Signal Transduction Pathways in Nitric Oxide-induced Tumor Apoptosis. Cancer Research. 61: 3682-3688, 2001. |23. Seshadri, V., et al. Dual role of insulin in transcriptional regulation of the acute phase reactant ceruloplasmin. JBC Papers in Press. Published on May 23, 2002 as Manuscript M203610200. |24. Treins, C. et al.Insulin stimulates HIF-1 through a PI-3-kinase/TOR-dependent signaling pathway. JBC Papers in Press. Published on May 24, 2002 as Manuscript M204152200. |25. Sutter, C.H, et al. Hypoxia-inducible factor 1 alpha protein expression is controlled by oxygen-regulated ubiquitination that is disrupted by deletions and missense mutations. PNAS. 97(9): 4748-4753, 2000. |26. Jogi, A., et al. Hypoxia alters gene expression in human neuroblastoma cells toward an immature and neural crest-like phenotype. PNAS. 98(4): 1583-1588, 2001. |27. Haase, V.H., et al. Vascular tumors in livers with targeted inactivation of the von Hippel-Lindau tumor suppressor. |PNAS. 98(4): 1583-1588, 2001. |28. Kamura, T., et al. Activation of HIF1alpha ubuitination by a reconstituted von Hippel-Lindau tumor suppressor complex. |PNAS. 97(19): 10430-10435, 2000. |29. Kim, M.S., et al. Histone deactylases induce angiogenesis by negative regulation of tumor suppressor genes. |Nature Medicine. 7: 437-443, 2001. |30. Palladino, M., et al. Expression of Hypoxia-Inducible Factor 1-alpha in the rat epidiymis. 35th Annual Meeting of the Society for the Study of Reproduction. 66(1): p.331 (abstract 577), 2002. |31. Fukuda, R., et al. IGF-1 Induces HIF-1-mediated VEGF Expression that is Dependent on MAP Kinase and PI-3-Kinase Signaling in Colon Cancer Cells. JBC Papers in Press. Published on July 30, 2002 as manuscript M203781200 |32. Palladino, M., et al. Stimulation of hypoxia-inducible factor-1 alpha protein in the adult rat testis following ischemic injury occurs without an increase in hif-1 alpha messenger RNA expression. Biology of Reproduction. 67: 995-1002, 2002 |33. Krieg, M., et al. Up-regulation of hypoxia-inducible factors HIF-1alpha and HIF-2alpha under normoxic conditions in renal carcinoma cells by von Hippel-Lindau tumor suppressor gene loss function. Oncogene. 19: 5435-5443, 2000 |34. Rapisarda, A., et al. Identification of Small Molecule Inhibitors of Hypoxia-Inducible Factor 1 Transcriptional Activation Pathway. Cancer Research. 62: 4316-4324, 2002 |||
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