HIF-2 alpha/EPAS1 Antibody Pack

Novus Biologicals | Catalog # NB100-902HIF2

Novus Biologicals
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Key Product Details

Species

Human, Mouse, Rat, Hamster, Rabbit

Applications

Chromatin Immunoprecipitation, ELISA, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunoprecipitation, Mycoplasma, Simple Western, Western Blot

Product Summary for HIF-2 alpha/EPAS1 Antibody Pack

This pack contains 1 vial each of: NB100-122SS (0.025 mL) and NB100-132SS (0.05 mL).

Product Specifications

Specificity

These antibodies are specific for HIF-2alpha.

Application Notes

See individual datasheets of components for their validated applications. By Western blot, these antibodies recognize a band ~118 kDa, representing HIF-2a in induced tissues and cells. Nuclear extracts should be used for Western analysis, if possible.

Reactivity Notes

See individual datasheets of components for their validated species

Scientific Data Images for HIF-2 alpha/EPAS1 Antibody Pack

Immunohistochemistry: HIF-2 alpha/EPAS1 Antibody Pack [NB100-902HIF2]

Immunohistochemistry: HIF-2 alpha/EPAS1 Antibody Pack [NB100-902HIF2]

Immunohistochemistry: HIF-2 alpha/EPAS1 Antibody Pack [NB100-902HIF2] - Analysis of HIF-2 alpha in human endometrium using. Donkey anti-mouse Alexa Fluor 488 secondary antibody was used. Image from verified customer review. HIF-2 alpha/EPAS1 Antibody (ep190b) [NB100-132]
Western Blot: HIF-2 alpha/EPAS1 Antibody Pack [NB100-902HIF2]

Western Blot: HIF-2 alpha/EPAS1 Antibody Pack [NB100-902HIF2]

Western Blot: HIF-2 alpha/EPAS1 Antibody Pack [NB100-902HIF2] - WNT11 is induced by hypoxia or hypoxic mimetics in different cell types. Immunoblot analyses of HeLa cells under normal air or hypoxia for 24 hrs. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep21520) licensed under a CC-BY license. HIF-2 alpha/EPAS1 Antibody [NB100-122]
Simple Western: HIF-2 alpha/EPAS1 Antibody Pack [NB100-902HIF2]

Simple Western: HIF-2 alpha/EPAS1 Antibody Pack [NB100-902HIF2]

Simple Western: HIF-2 alpha/EPAS1 Antibody Pack [NB100-902HIF2] - Lane view shows a specific band for HIF-2 alpha in 0.5 mg/ml of Hypoxic HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system. HIF-2 alpha/EPAS1 Antibody (ep190b) [NB100-132]
Knockout Validated: HIF-2 alpha/EPAS1 Antibody Pack [NB100-902HIF2]

Western Blot: HIF-2 alpha/EPAS1 Antibody Pack [NB100-902HIF2]

Western Blot: HIF-2 alpha/EPAS1 Antibody Pack [NB100-902HIF2] - HIF-1alpha is the predominant transcriptional regulator of WNT11 expression during hypoxia. EMSCs isolated from the indicated mouse genotypes were infected with lentivirus expressing GFP or Cre recombinase. Non-infected cells and GFP infected cells served as controls. Immunoblot analyses of EMSCs derived from the indicated genotypes treated with 0.1 mM DMOG for 24 hrs. Attenuated WNT11 expression in Hif-1alpha KO EMSCs (lenti-Cre infected Hif-1af/f). Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep21520) licensed under a CC-BY license. HIF-2 alpha/EPAS1 Antibody [NB100-122]
Immunohistochemistry: HIF-2 alpha/EPAS1 Antibody Pack [NB100-902HIF2]

Immunohistochemistry: HIF-2 alpha/EPAS1 Antibody Pack [NB100-902HIF2]

Immunohistochemistry: HIF-2 alpha/EPAS1 Antibody Pack [NB100-902HIF2] - Formalin-fixed paraffin-embedded tissue sections of human placenta were probed for HIF-2 alpha/EPAS1 mRNA (ACD RNAScope Probe, catalog #410598; Fast Red chromogen, ACD catalog # 322750). Adjacent tissue section was processed for immunohistochemistry using Rabbit Polyclonal (Novus Biologicals catalog # NB100-122) at 1:100 dilution with one-hour incubation at room temperature followed by incubation with anti-rabbit IgG VisUCyte HRP Polymer Antibody (Catalog # VC003) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue). Specific staining was localized to trophoblastic cells. HIF-2 alpha/EPAS1 Antibody [NB100-122]
Immunohistochemistry: HIF-2 alpha/EPAS1 Antibody Pack [NB100-902HIF2]

Immunohistochemistry: HIF-2 alpha/EPAS1 Antibody Pack [NB100-902HIF2]

Immunohistochemistry: HIF-2 alpha/EPAS1 Antibody Pack [NB100-902HIF2] - Normal ganglia 1 (NG1), normal ganglia 2 (NG2) and IDH-mutant (IDH). Sporadic PGL (Spo. PGL). HIF2 alpha staining. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0127471) licensed under a CC-BY license. HIF-2 alpha/EPAS1 Antibody (ep190b) [NB100-132]

Formulation, Preparation, and Storage

Concentration

Concentration of individual antibodies may be found on the vial label. If unlisted please contact technical services.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Storage

Storage is content dependent.

Background: HIF-2 alpha/EPAS1

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).

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. 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

Long Name

Hypoxia-inducible Transcription Factor 2 alpha

Alternate Names

EPAS1, HIF 2A, HIF2 alpha, HIF2A, HLF, MOP2

Gene Symbol

EPAS1

Additional HIF-2 alpha/EPAS1 Products

Product Documents for HIF-2 alpha/EPAS1 Antibody Pack

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Product Specific Notices for HIF-2 alpha/EPAS1 Antibody Pack

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.

Citations for HIF-2 alpha/EPAS1 Antibody Pack

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    Verified Customer | Posted 05/05/2010

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Protocols

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FAQs for HIF-2 alpha/EPAS1 Antibody Pack

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    • Q: 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?
      A: 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.

    • Q: I would like to know the working concentration for the 2 HIF-2 alpha antibodies in the kit NB100-902HIF2.
      A: 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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