HIF-2 alpha/EPAS1 Antibody - BSA Free

Images

 
Biological Strategies: 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 ...read more
Biological Strategies: 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 ...read more
Orthogonal Strategies: 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). ...read more
Expression of hypoxia-inducible alpha subunits in normal and diseased lung tissue. HIF-2alpha expression are more evident in fibroblasts from idiopathic pulmonary fibrosis (d) than from lung tissue affected by other ...read more
Image 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.
Genetic Strategies: Immunoblot validation of HIF2A and PHD2 KO clones using HIF2A (#NB100-122; dilution: 1/300), PHD2 (#NB100-137; dilution: 1/500). To blot HIFs factor cells were first pre-treated for 5 h with ...read more
Biological Strategies: HIF-2 alpha in human retinal and choroidal primary endothelia lysates using .. Image from verified customer review.
Biological Strategies: HIF-2 alpha in MDA-MB-231 cell lysate (overexpression and endogenous samples) using . did not react to HIF-1 alpha overexpression. Image from verified customer review.
Induced WNT11 expression with tumor hypoxia and WNT11 regulates tumor growth. Bevacizumab increased expression of HIF-1alpha and HIF-2alpha and WNT11. First 10 lanes are control tumors, and the last 10 lanes are tumors ...read more
MDA-MB-231 cells were exposed to 20% or 1% O2 for 16 hours, and chromatin immunoprecipitation (ChIP) was performed with the indicated antibody (Ab). Primers flanking the HIF binding site were used for qPCR. ChIP image ...read more
Biological Strategies: Lane 1: Cobalt chloride treated COS7 nuclear extracts. Lane 2: Untreated COS7 nuclear extracts.
786-O cells without or with VHL overexpression. Image from verified customer review.
Biological Strategies: Analysis using the HRP conjugate of NB100-122. Detection of normoxic and hypoxic nuclear rat cell lysates.
HIF-2 alpha immunoreactivity in human cardiac myocytes stained with NB100-122.
Expression and correlation of 3 antibodies: HIF-1 alpha, HIF-2 alpha and HAF in ccRCC. (A, C & E) Positive nuclear staining to the 3 antibodies: HIF-1 alpha, HIF-2 alpha and HAF, in primary ccRCCs at high magnification ...read more
Genetic Strategies: Reduction of HIF-2alpha levels leads to protection in UV-triggered apoptosis, but not for apoptosis caused by glucose and serum starvation in 786-O cells. Parental 786-O or those either stably ...read more

Product Details

Summary
Reactivity Hu, Mu, Rt, Fi, Ha, Pm, Rb, Re, ShSpecies Glossary
Applications WB, Simple Western, ELISA, Flow, Func, GS, ICC/IF, IHC, IP, PAGE, WB, ChIP, Dual ISH-IHC, KD, KO
Clonality
Polyclonal
Host
Rabbit
Conjugate
Unconjugated
Format
BSA Free
Concentration
1.0 mg/ml

Order Details

HIF-2 alpha/EPAS1 Antibody - BSA Free Summary

Immunogen
This HIF-2 alpha/EPAS1 Antibody was developed against a peptide derived from the C-terminus of mouse/human HIF-2 alpha protein.
Specificity
This HIF-2 alpha/EPAS1 Antibody is specific for HIF-2 alpha/EPAS, and does not cross-react with HIF-1 alpha.
Isotype
IgG
Clonality
Polyclonal
Host
Rabbit
Gene
EPAS1
Purity
Immunogen affinity purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Chromatin Immunoprecipitation (ChIP) 1:10-1:500
  • Dual RNAscope ISH-IHC
  • ELISA 1:100 - 1:2000
  • Flow Cytometry
  • 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-Frozen
  • 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)
  • SDS-Page
  • 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.
Simple Western reported by an internal validation. 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.
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.
Control
COS-7 Nuclear Hypoxic Induced Cell Lysate
Human Rectum Whole Tissue Lysate (Adult Whole Normal)
NIH 3T3 CoCl2 treated/Untreated Cell Lysate
L929 CoCl2 treated/Untreated Cell Lysate
Raw 264.7 CoCl2 treated/Untreated Cell Lysate
PC12 CoCl2 treated/Untreated Cell Lysate
PC12 Hypoxic / Normoxic Cell Lysate
Raw 264.7 Hypoxic / Normoxic Cell Lysate
L929 Hypoxic / Normoxic Cell Lysate
NIH 3T3 Hypoxic / Normoxic Cell Lysate
Reviewed Applications
Read 38 Reviews rated 4.4
using
NB100-122 in the following applications:

Publications
Read Publications using
NB100-122 in the following applications:

Reactivity Notes

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

Packaging, Storage & Formulations

Storage
Store at -20 °C.
Buffer
PBS
Preservative
0.05% Sodium Azide
Concentration
1.0 mg/ml
Purity
Immunogen affinity purified

Alternate Names for HIF-2 alpha/EPAS1 Antibody - BSA Free

  • anti hif 2 alpha
  • anti-HIF-2 alpha
  • Basic-helix-loop-helix-PAS protein MOP2
  • BHLHE73
  • Class E basic helix-loop-helix protein 73
  • ECYT4
  • endothelial PAS domain protein 1
  • endothelial PAS domain-containing protein 1
  • EPAS1
  • EPAS-1
  • HIF 2A
  • HIF-1-alpha-like factor
  • HIF-1alpha-like factor
  • HIF2 alpha
  • HIF-2 alpha
  • hif2a angiogenesis
  • HIF2A
  • HIF-2-alpha
  • HIF2-alpha
  • HLF
  • hypoxia-inducible factor 2 alpha
  • Hypoxia-inducible factor 2-alpha
  • Member of PAS protein 2
  • MOP2
  • PAS domain-containing protein 2
  • PASD2

Background

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

Limitations

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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Publications for HIF-2 alpha/EPAS1 Antibody (NB100-122)(765)

We have publications tested in 9 confirmed species: Human, Mouse, Rat, Golden Syrian Hamster, Hamster, Rabbit, Reptile, Sheep, Zebrafish.

We have publications tested in 21 applications: B/N, ChIP, Chemotaxis, Chip Cytometry, CoIP, ELISA, FLOW, Func, GS, IB, ICC, ICC/IF, IF/IHC, IHC-Fr, IHC-P, IP, In Vivo, KD, KO, SW, WB.


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(3)
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(21)
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(31)
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(39)
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Showing Publications 1 - 10 of 765. Show All 765 Publications.
Publications using NB100-122 Applications Species
Liao Q, Zhu C, Sun X et al. Disruption of sirtuin 7 in zebrafish facilitates hypoxia tolerance Journal of Biological Chemistry 2023-08-01 [PMID: 37481210] (In Vivo, B/N) In Vivo, B/N
Kubaichuk K, Kietzmann T USP10 Contributes to Colon Carcinogenesis via mTOR/S6K Mediated HIF-1? but Not HIF-2? Protein Synthesis Cells 2023-06-08 [PMID: 37371055] (WB, Human, Mouse) WB Human, Mouse
Yue Y, Tang Y, Huang H et al. VBP1 negatively regulates CHIP and selectively inhibits the activity of hypoxia-inducible factor (HIF)-1? but not HIF-2? The Journal of biological chemistry 2023-05-16 [PMID: 37201586] (WB, Zebrafish) WB Zebrafish
Aboouf MA, Armbruster J, Guscetti F et al. Endogenous myoglobin expression in mouse models of mammary carcinoma reduces hypoxia and metastasis in PyMT mice Scientific reports 2023-05-09 [PMID: 37161046] (IHC-P, Mouse) IHC-P Mouse
Qu M, Long Y, Wang Y et al. Hypoxia Increases ATX Expression by Histone Crotonylation in a HIF-2?-Dependent Manner International Journal of Molecular Sciences 2023-04-11 [PMID: 37108194] (B/N) B/N
Leu T, Denda J, Wrobeln A, Fandrey J Hypoxia-Inducible Factor-2alpha Affects the MEK/ERK Signaling Pathway via Primary Cilia in Connection with the Intraflagellar Transport Protein 88 Homolog Molecular and cellular biology 2023-04-19 [PMID: 37074220]
Li KQ, Liu GJ, Liu XY et al. EPAS1 prevents telomeric damage-induced senescence by enhancing transcription of TRF1, TRF2, and RAD50 Zoological Research 2023-05-18 [PMID: 37070589] (WB, ICC/IF) WB, ICC/IF
Sun H, Pratt RE, Dzau VJ, Hodgkinson CP Neonatal and adult cardiac fibroblasts exhibit inherent differences in cardiac regenerative capacity The Journal of biological chemistry 2023-04-10 [PMID: 37044217] (WB, ICC/IF, Mouse) WB, ICC/IF Mouse
Mayro B, Hoj JP, Cerda-Smith CG et al. ABL kinases regulate the stabilization of HIF-1? and MYC through CPSF1 Proceedings of the National Academy of Sciences of the United States of America 2023-04-18 [PMID: 37040401]
Ye J, Chen J, Li Y et al. Hepatocyte-specific knockout of HIF-2? cannot alleviate carbon tetrachloride-induced liver fibrosis in mice PeerJ 2023-04-03 [PMID: 37033734] (IHC-P, Mouse) IHC-P Mouse