Porcine HIF-1 alpha ELISA Kit (Colorimetric)
Novus Biologicals, part of Bio-Techne | Catalog # NBP3-42577
Key Product Details
Sample Type & Volume Required Per Well
Serum, Plasma, Tissue homogenates and other biological fluids (100 uL)
Sensitivity
6.4 pg/mL
Assay Range
15.6 - 1000 pg/mL
Product Specifications
Assay Type
Sandwich ELISA
Kit Type
ELISA Kit (Colorimetric)
Reactivity
Porcine
Description
Assay time: 3 hours
Precision
Intra-Assay Precision (Precision within an assay) %CV < 10
Inter-Assay Precision (Precision between assays) %CV < 12
Recovery for Porcine HIF-1 alpha ELISA Kit (Colorimetric)
Recovery
83 - 97%
Kit Contents for Porcine HIF-1 alpha ELISA Kit (Colorimetric)
- Assay Diluent A
- Assay Diluent B
- Detection Reagent A
- Detection Reagent B
- Instruction manual
- Plate sealer for 96 wells
- Pre-coated ready to use 96-well strip plate
- Standard
- Standard Diluent
- Stop Solution
- TMB Substrate
- Wash Buffer (30 x concentrate)
Preparation and Storage
Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Storage of components varies. See protocol for specific instructions.
Background: HIF-1 alpha/HIF1A
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
Long Name
Hypoxia Inducible Factor 1 Subunit Alpha
Alternate Names
BHLHE78, HIF 1A, HIF-1a, HIF1 alpha, HIF1A, MOP1, PASD8
Gene Symbol
HIF1A
Additional HIF-1 alpha/HIF1A Products
Product Documents for Porcine HIF-1 alpha ELISA Kit (Colorimetric)
Product Specific Notices for Porcine HIF-1 alpha ELISA Kit (Colorimetric)
This product is for research use only and is not approved for use in humans or in clinical diagnosis. ELISA Kits are guaranteed for 6 months from date of receipt.
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