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HIF-2 alpha/EPAS1 Antibody - BSA Free Best Seller

Novus Biologicals, part of Bio-Techne | Catalog # NB100-122

Novus Biologicals, part of Bio-Techne
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NB100-122
NB100-122SS

Key Product Details

Validated by

Knockout/Knockdown, Orthogonal Validation, Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat, Fish, Hamster, Primate, Rabbit, Reptile, Sheep

Cited:

Human, Mouse, Rat, Fish - Danio rerio (Zebrafish), Hamster, Ovine, Rabbit, Reptile

Applications

Validated:

Chromatin Immunoprecipitation (ChIP), Dual RNAscope ISH-IHC, ELISA, Flow Cytometry, Gel Super Shift Assays, Immunoblotting, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Immunoprecipitation, In vitro assay, Knockdown Validated, Knockout Validated, SDS-Page, Simple Western, Western Blot

Cited:

Block/Neutralize, Chemotaxis, Chip Cytometry, Chromatin Immunoprecipitation (ChIP), CoIP, ELISA, Flow Cytometry, Functional Assay, Gel Supershift Assay, IF/IHC, Immunoblotting, Immunocytochemistry, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Immunoprecipitation, In vivo assay, Knockdown Validated, Knockout Validated, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free

Concentration

1.0 mg/ml

Product Specifications

Immunogen

This HIF-2 alpha/EPAS1 Antibody was developed against a peptide derived from the C-terminus of mouse/human HIF-2 alpha protein.

Reactivity Notes

Use in Mouse reported in scientific literature (PMID:33758176).

Specificity

This HIF-2 alpha/EPAS1 Antibody is specific for HIF-2 alpha/EPAS, and does not cross-react with HIF-1 alpha.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

96.5 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for HIF-2 alpha/EPAS1 Antibody - BSA Free

Western Blot Analysis of HIF-2 alpha/EPAS1 in Hypoxic and non-Hypoxic HeLa Cells

Western Blot Analysis of HIF-2 alpha/EPAS1 in Hypoxic and non-Hypoxic HeLa Cells

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 (https://www.nature.com/articles/srep21520) licensed under a CC-BY license.
Knockout Validation of HIF-2 alpha/EPAS1 Antibody in Control and HIF-2 alpha Knockout Mouse EMSCs by Western Blot

Knockout Validation of HIF-2 alpha/EPAS1 Antibody in Control and HIF-2 alpha Knockout Mouse EMSCs by Western Blot

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 (https://www.nature.com/articles/srep21520) licensed under a CC-BY license.
Dual RNAscope ISH-IHC Analysis of HIF-2 alpha/EPAS1 in Human Placenta

Dual RNAscope ISH-IHC Analysis of HIF-2 alpha/EPAS1 in Human Placenta

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.

Applications for HIF-2 alpha/EPAS1 Antibody - BSA Free

Application
Recommended Usage

Chromatin Immunoprecipitation (ChIP)

1:10-1:500

ELISA

1:100 - 1:2000

Gel Super Shift Assays

reported in scientific literature (PMID 15184875)

Immunoblotting

reported in scientific literature (PMID 28115701)

Immunocytochemistry/ Immunofluorescence

1:100

Immunohistochemistry

1:100

Immunohistochemistry-Paraffin

1:100

Immunoprecipitation

5 ug / 1 mg lysate

In vitro assay

reported in scientific literature (PMID 24998849)

Knockdown Validated

reported in scientific literature (PMID 31061092)

Knockout Validated

reported in scientific literature (PMID 26861754)

Simple Western

1:50

Western Blot

1 - 2 ug/mL
Application Notes
In WB, this antibody recognizes a band at 118 kDa representing HIF-2 alpha.
See Simple Western Antibody Database for Simple Western validation: tested in hypoxic HeLa lysate (0.5 mg/ml); separated by Size- Wes/Sally Sue/Peggy Sue; antibody dilution of 1:50; apparent MW in kDa on Simple Western was 110kDa; matrix was 12-230 kDa.

Reviewed Applications

Read 39 reviews rated 4.4 using NB100-122 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.05% Sodium Azide

Concentration

1.0 mg/ml

Shipping

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

Stability & Storage

Store at -20 °C.

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, anti hif 2 alpha, anti-HIF-2 alpha

Gene Symbol

EPAS1

Additional HIF-2 alpha/EPAS1 Products

Product Documents for HIF-2 alpha/EPAS1 Antibody - BSA Free

Certificate of Analysis

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

Product Specific Notices for HIF-2 alpha/EPAS1 Antibody - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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