Key Product Details

Validated by

Biological Validation

Species Reactivity

Human, Mouse, Rat, Bovine, Hamster

Applications

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, ELISA, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Simple Western, Immunoprecipitation, Chromatin Immunoprecipitation (ChIP), In vivo assay, Gel Super Shift Assays, Gel Supershift Assay, Knockdown Validated, CyTOF-ready

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # ep190b

Format

Azide and BSA Free
View all HIF-2 alpha/EPAS1 Primary Antibodies »

Product Specifications

Immunogen

The immunogen recognized by this HIF-2 alpha/EPAS1 Antibody (ep190b) maps to a region between amino acids 535-631. [UniProt# Q99814]

Reactivity Notes

Ability to use HIF-2 alpha/EPAS1 Antibody (ep190b) in mouse is mixed with some positive and some negative results.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

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 (ep190b) - Azide and BSA Free

Western Blot: HIF-2 alpha/EPAS1 Antibody (ep190b)Azide and BSA Free [NBP2-80766]

Western Blot: HIF-2 alpha/EPAS1 Antibody (ep190b)Azide and BSA Free [NBP2-80766]

Western Blot: HIF-2 alpha/EPAS1 Antibody (ep190b) - Azide and BSA Free [NBP2-80766] - Western blot analysis and quantification of HIF-2 alpha expression in the cortex 4.5 hours after lipopolysaccharide (LPS) administration for all LPS groups and the control group (SHAM, white bar). With the exception of the vagotomy group (VGX LPS, gray bar), no significant differences to the SHAM group were found in the LPS-treated and sham-operated (SHAM+LPS, light gray bar) or vagus nerve-stimulated groups (VGX LPS+STIM, black bar). The significant increase in the VGX LPS (gray bar) group is an indicator of a hypoxic condition; * P<0.05 compared to SHAM; n=6 rats each. Data are given as the mean+/-SEM. Image collected and cropped by CiteAb from the following publication (https://jneuroinflammation.biomedcentral.com/articles/10.1186/1742-2094…), licensed under a CC-BY license. Image from the standard format of this antibody.
Immunohistochemistry: HIF-2 alpha/EPAS1 Antibody (ep190b) - Azide and BSA Free [NBP2-80766]

Immunohistochemistry: HIF-2 alpha/EPAS1 Antibody (ep190b) - Azide and BSA Free [NBP2-80766]

Immunohistochemistry: HIF-2 alpha/EPAS1 Antibody (ep190b) - Azide and BSA Free [NBP2-80766] - Renal tubule specific models of Vhl deletion. Histological images of representative renal sections from 12 month old control, Pax8-CreERT2/Vhldelta/delta and Slc22a6-CreERT2/Vhldelta/delta mice (stains and antibodies as indicated, arrowheads indicate abno
Flow Cytometry: HIF-2 alpha/EPAS1 Antibody (ep190b) - Azide and BSA Free [NBP2-80766]

Flow Cytometry: HIF-2 alpha/EPAS1 Antibody (ep190b) - Azide and BSA Free [NBP2-80766]

Flow Cytometry: HIF-2 alpha/EPAS1 Antibody (ep190b) - Azide and BSA Free [NBP2-80766] - HIF-2 alpha antibody was tested at 1:400 in HepG2 cells using an Alexa Fluor 488 secondary (shown in purple). M1 is defined by unstained cells. Image from the standard format of this antibody.
Western Blot: HIF-2 alpha/EPAS1 Antibody (ep190b)Azide and BSA Free [NBP2-80766]

Western Blot: HIF-2 alpha/EPAS1 Antibody (ep190b)Azide and BSA Free [NBP2-80766]

Western Blot: HIF-2 alpha/EPAS1 Antibody (ep190b) - Azide and BSA Free [NBP2-80766] - 4-OOHCPA exposure induced HIF-2 alpha/EPAS1. A: Western blot analysis of HIF-2 alpha/EPAS1 protein (118KD) and actin (43KD) in limbs at 10 min, 1, 3, 6, and 24 h after treatment with 4-OOHCPA at 0.3 ug/mL (L) 1.0 ug/mL (M) or 3.0 ug/mL (H). P represents the positive control. Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0051937), licensed under a CC-BY license. Image from the standard format of this antibody.
Western Blot: HIF-2 alpha/EPAS1 Antibody (ep190b)Azide and BSA Free [NBP2-80766]

Western Blot: HIF-2 alpha/EPAS1 Antibody (ep190b)Azide and BSA Free [NBP2-80766]

Western Blot: HIF-2 alpha/EPAS1 Antibody (ep190b) - Azide and BSA Free [NBP2-80766] - Analysis of HepG2 without Cobalt (II) Chloride (1), HepG2 with Cobalt (II) Chloride (2), HepG2 normoxic (3), HepG2 hypoxic (4), HepG2 without Cobalt (II) Chloride (5), HepG2 with Cobalt (II) Chloride (6), HepG2 normoxic (7), and HepG2 hypoxic (8) using this antibody (NB100-132) at 1 - 2 ug/mL. Image from the standard format of this antibody.
Western Blot: HIF-2 alpha/EPAS1 Antibody (ep190b)Azide and BSA Free [NBP2-80766]

Western Blot: HIF-2 alpha/EPAS1 Antibody (ep190b)Azide and BSA Free [NBP2-80766]

Western Blot: HIF-2 alpha/EPAS1 Antibody (ep190b) - Azide and BSA Free [NBP2-80766] - Analysis of HIF-2 alpha stabilization over time in 791T cells following exposure to hypoxia. [PMID: 23785417]. Image using the HRP form of this antibody. Image from the standard format of this antibody.
Western Blot: HIF-2 alpha/EPAS1 Antibody (ep190b)Azide and BSA Free [NBP2-80766]

Western Blot: HIF-2 alpha/EPAS1 Antibody (ep190b)Azide and BSA Free [NBP2-80766]

Western Blot: HIF-2 alpha/EPAS1 Antibody (ep190b) - Azide and BSA Free [NBP2-80766] - Analysis of HIF-2 alpha stabilization over time in HOS cells following exposure to hypoxia. [PMID: 23785417]. Image using the HRP form of this antibody. Image from the standard format of this antibody.
Western Blot: HIF-2 alpha/EPAS1 Antibody (ep190b)Azide and BSA Free [NBP2-80766]

Western Blot: HIF-2 alpha/EPAS1 Antibody (ep190b)Azide and BSA Free [NBP2-80766]

Western Blot: HIF-2 alpha/EPAS1 Antibody (ep190b) - Azide and BSA Free [NBP2-80766] - Mouse aortic endothelial cells treated (1%) or not treated (20.9%) in hypoxia for 3 hrs. Cells where also transfected with a specific siRNA against (siHIF-2) or a control siRNA (-). Western blot image submitted by a verified customer review. Image from the standard format of this antibody.
Immunohistochemistry: HIF-2 alpha/EPAS1 Antibody (ep190b) - Azide and BSA Free [NBP2-80766]

Immunohistochemistry: HIF-2 alpha/EPAS1 Antibody (ep190b) - Azide and BSA Free [NBP2-80766]

Immunohistochemistry: HIF-2 alpha/EPAS1 Antibody (ep190b) - Azide and BSA Free [NBP2-80766] - Analysis of HIF-2 alpha in human endometrium using. Donkey anti-mouse Alexa Fluor 488 secondary antibody was used. IHC image submitted by a verified customer review. Image from the standard format of this antibody.
Immunohistochemistry: HIF-2 alpha/EPAS1 Antibody (ep190b) - Azide and BSA Free [NBP2-80766]

Immunohistochemistry: HIF-2 alpha/EPAS1 Antibody (ep190b) - Azide and BSA Free [NBP2-80766]

Immunohistochemistry: HIF-2 alpha/EPAS1 Antibody (ep190b) - Azide and BSA Free [NBP2-80766] - Analysis of HIF-2 in human cardiac myocytes using HIF-2 alpha/EPAS1 Antibody (ep190b). Image from the standard format of this antibody.
Simple Western: HIF-2 alpha/EPAS1 Antibody (ep190b)Azide and BSA Free [NBP2-80766]

Simple Western: HIF-2 alpha/EPAS1 Antibody (ep190b)Azide and BSA Free [NBP2-80766]

Simple Western: HIF-2 alpha/EPAS1 Antibody (ep190b) - Azide and BSA Free [NBP2-80766] - 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. Image from the standard format of this antibody.

Applications for HIF-2 alpha/EPAS1 Antibody (ep190b) - Azide and BSA Free

Application
Recommended Usage

ELISA

1:100-1:2000

Flow Cytometry

1:400

Gel Super Shift Assays

reported in scientific literature (PMID 17404621)

Immunohistochemistry

1:150 - 1:300

Immunohistochemistry-Frozen

reported in scientific literature (PMID 24973414)

Immunohistochemistry-Paraffin

1:150 - 1:300

In vivo assay

reported in scientific literature (PMID 23857308)

Simple Western

1:100

Western Blot

1 - 2 ug/mL
Application Notes
In WB, this HIF-2 alpha (ep190b) antibody recognizes a band at approx. 118 kDa representing HIF-2 alpha.

In Simple Western only 10 - 15 uL of the recommended dilution is used per data point.
See Simple Western Antibody Database for Simple Western validation: antibody dilution of 1:100. Separated by Size-Wes, Sally Sue/Peggy Sue.
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.

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS

Format

Azide and BSA Free

Preservative

No Preservative

Concentration

1 mg/ml

Shipping

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

Stability & Storage

Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.

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 (ep190b) - Azide and BSA Free

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Product Specific Notices for HIF-2 alpha/EPAS1 Antibody (ep190b) - Azide and 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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Protocols

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