A 96-well assay for measurement of HIF-1α
Features
  • A sensitive, non-radioactive method for detecting HIF-1α from whole cell lysates
  • 96-well plate format replaces EMSAs
  • Capture the transcription factor using a specific dsDNA sequence bound to the plate
  • Detect the dsDNA-bound transcription factor with specific antibodies in an ELISA format
  • Nuclear Extraction Kit (Item No. 10009277) available to aid in the isolation of nuclear and cytoplasmic fractions from cell lysates or tissue homogenates
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HIF-1α Transcription Factor Assay Kit

Item No. 10006910

Kit Booklet (PDF)Safety Data Sheet (SDS) (PDF)
Technical Information
Synonyms
  • ARNT-interacting Protein
  • Hypoxia-Inducible Factor-1α
Origin
Animal/Bovine, Animal/Goat, Animal/Rabbit
Shipping & Storage Information
Storage
-80°C
Shipping
Dry ice in continental US; may vary elsewhere
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Certificates of Analysis & Batch Specific Data

Provide batch numbers separated by commas to download or request available product inserts, QC sheets, certificates of analysis, data packs, and GC-MS data.

    96 wells

    $543

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    Product Description

    The HIF (hypoxia-inducible factor) transcription factor is a member of the basic-helix-loop-helix (bHLH) family of transcription factors and plays an important role in maintaining cellular oxygen homeostasis.1,2 HIF-1α has emerged as an important drug target in breast and prostate cancer, cardiovascular disease, and ischemia.3,4,5 Cayman’s HIF-1α Transcription Factor Assay is a non-radioactive, sensitive method for detecting specific transcription factor DNA binding activity in nuclear extracts and whole cell lysate. A 96-well enzyme-linked immunosorbent assay (ELISA) replaces the cumbersome radioactive electrophoretic mobility shift assay (EMSA). A specific double stranded DNA (dsDNA) consensus sequence containing the HIF-1α response element is immobilized to the wells of a 96-well plate. HIF-1α contained in a nuclear extract or whole cell lysates, binds specifically to the HIF-1α response element. The HIF transcription factor complex is detected by addition of a specific primary antibody directed against HIF-1α. A secondary antibody conjugated to HRP is added to provide a sensitive colorimetric readout at 450 nm.

    Needed but not supplied: Please download the kit booklet to verify if UltraPure Water (Milli-Q or equivalent) or any other components are needed for this assay.

    WARNING This product is not for human or veterinary use.

    References & Product Citations
    Product Description References

    1. Wang, G.L., Jiang, B.H., Rue, E.A., et al. Hypoxia-inducible factor 1 is a basic-helix-loop-helix-PAS heterodimer regulated by cellular O2 tension. Proc. Natl. Acad. Sci. USA 92(12), 5510-5514 (1995).

    2. Safran, M., and Kaelin, W.G., Jr. HIF hydroxylation and the mammalian oxygen-sensing pathway. J. Clin. Invest. 111(6), 779-783 (2003).

    3. Rapisarda, A., Uranchimeg, B., Scudiero, D.A., et al. Identification of small molecule inhibitors of hypoxia-inducible factor 1 transcriptional activation pathway. Cancer Res. 62, 4316-4324 (2002).

    4. William, C., Masson, N., Tian, Y.M., et al. Peptide blockade of HIFα degradation modulates cellular metabolism and angiogenesis. Proc. Natl. Acad. Sci. USA 99(16), 10423-10428 (2002).

    5. Welsh, S., Williams, R., Kirkpatrick, L., et al. Antitumor activity and pharmacodynamic properties of PX-478, an inhibitor of hypoxia-inducible factor-1α. Mol. Cancer Ther. 3(3), 233-244 (2004).

    Product Citations

    Jain, M., Joharapurkar, A., Patel, V., et al. Pharmacological inhibition of prolyl hydroxylase protects against inflammation-induced anemia via efficient erythropoiesis and hepcidin downregulation. Eur. J. Pharmacol. 843, 113-120 (2019).

    Jain, M.R., Joharapurkar, A.A., Pandya, V., et al. Pharmacological characterization of ZYAN1, a novel prolyl hydroxylase inhibitor for the treatment of anemia. Drug Res. (Stuttg.) 66(2), 107-112 (2016).

    Yang, L., Xie, M., Yang, M., et al. PKM2 regulates the Warburg effect and promotes HMGB1 release in sepsi. Nat. Commun. 5, 4436 (2014).