Hypoxia Inducible Factor 1a (HIF-1a)

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 that activates gene transcription in response to reduced cellular O2 concentration. HIF-1 activates the transcription of EPO, VEGF, iNOS, heme oxygenase 1 and several other critical intracellular responses to hypoxia. HIF-1 is a heterodimer composed of HIF-1a and HIF-1b subunits. Under normoxic conditions, human HIF-1a is hydroxylated at Pro402 or Pro564, is polyubiquinated, and eventually degraded. Under hypoxic conditions, the lack of hydroxylation prevents HIF degradation and increases transcriptional activity. Therefore, the concentration of HIF-1a increases in the cell. HIF-1b remains stable under either condition. HIF hydroxylases provide insight into hypoxic cell responses, which may be used to help isolate therapeutic agents.\n\nApplications: \nSuitable for use in Western Blot, Immunohistochemistry and Immunoprecipitation. Other applications not tested.\n\nRecommended Dilution:\nWestern Blot: 1:250 (ECL, nuclear extracts). Recognizes a band at 120kD representing HIF-1 alpha in induced tissues and cells. Multiple bands may be seen representing post-translational modification. The mobility of HIF-1 alpha induced by desferrioxamine or cobalt chloride treatment differs from the mobility of the hypoxia-induced protein. The reason is not known. \nImmunohistochemistry (paraffin tissues): 1:250-1:500 \nOptimal dilutions to be determined by the researcher.\n\nPositive Control:\nWestern Blot: COS-7 treated and untreated nuclear extracts can be used for positive and negative controls for hypoxic upregulation.\nImmunohistochemistry: Glioblastoma multiforme.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage, store at -20 degrees C. Aliquots are stable for at least 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

H9807-02

Size

100ul

Applications

IHC, IP, WB

Hosts

Mouse

Reactivities

Hum, Mouse, Rat, Bov, NHP, Prc, Sheep

Form

Supplied as a liquid in PBS, pH 7.2, 0.05% sodium azide, before the addition of glycerol to 40%.

P Type

Mab

Purity

Purified by Protein G affinity chromatography.

Isotype

IgG2b

References

1. Zhong, H., et al., Can. Res. 59: 5830-5835 (1999). 2. Duyndam, M.C.A., et al., J. Biol. Chem. 276(51): 48,066-48,076 (2001). 3. An, W.G., et al., Genes & Devel. 12: 149-162 (1998). 4. Iyer, N.V., et al., Genes & Devel. 12: 149-162 (1998). 5. Kaelin, W.G., Nature 399: 203-204 (1999). 6. Lando, D., et al., Genes & Devel. 16: 1466-1471 (2002). 7. Ozaki, H., et al., Inves. Ophthalmology & Visual Sci. 40(1): 182-189 (1999). 8. Palmer, L.A., et al., Amer. Physiol. Soc. 274: L212-L219 (1998). 9. Semenza, G.L., J. Lab. Clin. 131: 207-214 (1998). 10. Semenza, G.L., Genes & Devel. 14: 1983-1991 (2000). 11. Semenza, G.L., Biocem. Pharm. 59: 47-52 (2000). 12. Takahata, S., et al., Biochem. & Biophys. Res. Comm. 248: 789-794 (1998). 13. Wang, G.L. & Semenza, G.L., J. Biol. Chem. 270(3): 1230-1237 (1995). 14. Wang, G.L., et al., PNAS 92: 5510-5514 (1995). 15. de Candia, P., et al., PNAS 100(21): 12,337-12,342 (2003). 16. Fujii, T., et al., Int. J. Oncology 20: 955-962 (2002). 17. Mandiota, S.J., et al., Cancer Cell 1: 459-468 (2002). 18. Tang, T., et al., J. Biol. Chem. 277(21): 18,365-18,372 (2002). 19. Burke, B., et al., Amer. J. of Pathology 163(4): 1233-1242 (2003). 20. Franco, M., et al., Can. Res. 66: 7 (2006).

Additional Info

Recognizes human HIF-1 alpha at 120kD. Species Crossreactivity: monkey, sheep, mouse, rat, porcine, bovine and ferret.