HIF-2 alpha/EPAS1 Antibody Pack Summary
For NB100-122: Proprietary peptide derived from the mouse/human HIF-2a. For NB100-132: Human HIF-2a amino acids 535-631.
These antibodies are specific for HIF-2alpha.
- Chromatin Immunoprecipitation (ChIP)
- Flow Cytometry
- Knockout Validated
- Simple Western
- Western Blot
By Western blot, these antibodies recognize a band ~118 kDa, representing HIF-2a in induced tissues and cells. The monoclonal has been used for immunohistochemistry. The polyclonal has also been used, but success has varied. Nuclear extracts should be used for Western analysis, if possible.
See individual datasheets for validated applications.
NB 100-122 recognizes human, mouse, and rat HIF-2a. Other species have not been tested.NB 100-132 reacts with human and rat HIF-2 alpha. Results in mouse have been mixed (some + and some -). Other species have not been tested.
Packaging, Storage & Formulations
Storage is content dependent.
This pack contains 1 vial each of: NB100-122SS (0.025 mL) and NB100-132SS (0.05 mL).
Alternate Names for HIF-2 alpha/EPAS1 Antibody Pack
- Basic-helix-loop-helix-PAS protein MOP2
- Class E basic helix-loop-helix protein 73
- endothelial PAS domain protein 1
- endothelial PAS domain-containing protein 1
- HIF 2A
- HIF-1-alpha-like factor
- HIF-1alpha-like factor
- HIF2 alpha
- HIF-2 alpha
- hif2a angiogenesis
- hypoxia-inducible factor 2 alpha
- Hypoxia-inducible factor 2-alpha
- Member of PAS protein 2
- PAS domain-containing protein 2
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, 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. Similarly, HIF-2 alpha plays a role in cellular responses to hypoxia, but whereas HIF-1 alpha is ubiquitously expressed, HIF-2 alpha is predominantly expressed in the vascular endothelium at embryonic stages and after birth in select cells and tissue types (e.g., fibroblasts, hepatocytes and myocytes at 96kDa) (4). Following a similar mechanism to HIF-1 alpha, HIF-2 alpha is stabilized under hypoxic conditions by the formation of a heterodimer with an ARNT/HIF-1 beta subunit. Stable HIF-2 alpha-ARNT/HIF-1 beta heterodimers engage p300/CBP in the nucleus for binding to hypoxic response elements (HREs), inducing transcription, and thus regulation of genes (e.g., EPO, VEGFA). HIF-1 predominantly transactivates genes involved in glycolytic control and pro-apoptotic genes (e.g., LDHA and BNIP3), and HIF-2 regulates the expression of genes involved in invasion and stemness (e.g., MMP2, and OCT4). Common gene targets for HIF-1 and HIF-2 include VEGFA and GLUT1 (5).
The HIF-2 alpha subunit is rapidly targeted and degraded by the ubiquitin proteasome system under normoxic conditions. This process is mediated by oxygen-sensing enzymes, prolyl hydroxylase domain enzymes (PHDs), which catalyze the hydroxylation of key proline residues (Pro-405 and Pro-531) within the oxygen-dependent degradation domain of HIF-2 alpha (5). Once hydroxylated, HIF-2 alpha binds the von Hippel-Lindau tumor suppressor protein (pVHL) for subsequent ubiquitination and proteasomal degradation (5,6).
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. Hu, C.-J., Wang, L.-Y., Chodosh, L. A., Keith, B., & Simon, M. C. (2003). Differential Roles of Hypoxia-Inducible Factor 1 (HIF-1) and HIF-2 in Hypoxic Gene Regulation. Molecular and Cellular Biology. https://doi.org/10.1128/mcb.23.24.9361-9374.2003
5. Koh, M. Y., & Powis, G. (2012). Passing the baton: The HIF switch. Trends in Biochemical Sciences. https://doi.org/10.1016/j.tibs.2012.06.004
6. 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
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Antibody Packs are guaranteed
for 1 year from date of receipt.
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Product General Protocols
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FAQs for HIF-2 alpha/EPAS1 Antibody Pack (NB100-902HIF2). (Showing 1 - 2 of 2 FAQs).
I am planning to order the hif2a antibody pack NB100-902HIF2. Do you know if the antibodies can be re-used? Is a 1:500 dilution recommended for ecl detection? Will the antibodies work in total cell extracts?
- We do not recommend re-using the antibodies and we will not guarantee their function if they are re-used. You can use a normal dilution of secondary antibody for use with these antibodies. We have only tested their use on whole cell lysates. Since HIF2a is a nuclear protein, you will want to make sure it will still be in your cell extracts.
I would like to know the working concentration for the 2 HIF-2 alpha antibodies in the kit NB100-902HIF2.
- The working dilutions for Western Blots are: NB100-122 we recommend a range of 1:200-1:1000 depending on your samples, and NB100-132 we recommend 1:500. They both come at a starting concentration of 1.0 mg/mL. If you have questions on other applications let us know. Additionally, all of the dilutions can be found on the respective datasheets should you need a quicker answer for the working concentration.
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