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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
Sp1 and HIFs contribute to the synergistic activation of multiple genes in OVSAYO cells under SSH.b) Activation of multiple genes under hypoxia is dependent on HIFs. Western blotting is also shown for HIFs. Data shown ...read more
Western Blot: Rabbit Polyclonal HIF-2 alpha/EPAS1 Antibody [NB100-122] - Analysis of HIF-2 alpha/EPAS1 antibody on mouse liver nuclear extracts. Image from a verified customer review.

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

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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.
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.
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 39 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)(822)

We have publications tested in 9 confirmed species: Human, Mouse, Rat, Bat, 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, IHC-Fr, IHC-P, IP, In Vivo, KD, KO, WB.


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(5)
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(2)
GS
(3)
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(21)
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(1)
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(32)
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(39)
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(5)
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(8)
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(20)
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(3)
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Showing Publications 1 - 10 of 822. Show All 822 Publications.
Publications using NB100-122 Applications Species
Siyong Peng, Yingbo Guo, Marie Irondelle, Abigail Mazzu, Michel Kahi, Paula Ferreira Montenegro, Frédéric Bost, Nathalie M. Mazure The marine-derived HIF-1 alpha inhibitor, Yardenone 2, reduces prostate cancer cell proliferation by targeting HIF-1 target genes Cellular & Molecular Biology Letters 2024-07-08 [PMID: 38978018]
Kenneth K. Y. Ting, Pei Yu, Riley Dow, Hisham Ibrahim, Saraf Karim, Chanele K. Polenz, Daniel A. Winer, Minna Woo, Jenny Jongstra-Bilen, Myron I. Cybulsky Cholesterol accumulation impairs HIF-1 alpha -dependent immunometabolic reprogramming of LPS-stimulated macrophages by upregulating the NRF2 pathway Scientific Reports 2024-05-15 [PMID: 38750095]
Chan D.K. Nguyen, Benjamín A. Colón-Emeric, Shigekazu Murakami, Mia N.Y. Shujath, Chunling Yi PRMT1 promotes epigenetic reprogramming associated with acquired chemoresistance in pancreatic cancer Cell reports 2024-06-28 [PMID: 38691454]
Sebastian Billig, Marc Hein, Celine Kirchner, David Schumacher, Moriz Aljoscha Habigt, Mare Mechelinck, Dieter Fuchs, Uwe Klinge, Alexander Thei beta en, Christian Beckers, Christian Bleilevens, Rafael Kramann, Moritz Uhlig Coronary Microvascular Dysfunction in Acute Cholestasis-Induced Liver Injury. Biomedicines 2024-04-16 [PMID: 38672230]
Milankumar Prajapati, Jared Z. Zhang, Lauren Chiu, Grace S. Chong, Courtney J. Mercadante, Heather L. Kowalski, Bradley Delaney, Jessica A. Anderson, Shuling Guo, Mariam Aghajan, Thomas B. Bartnikas Hepatic HIF2 is a key determinant of manganese excess and polycythemia in SLC30A10 deficiency JCI Insight 2024-04-23 [PMID: 38652538]
Jacinta H. Martin, Ilana R. Bernstein, Jess M. Lyons, Ariel R. Brady, Nishani S. Mabotuwana, Simone J. Stanger, Camila Salum De Oliveira, Katerina B. Damyanova, Brett Nixon, Tessa Lord EPAS1 expression contributes to maintenance of the primordial follicle pool in the mouse ovary Scientific Reports 2024-04-16 [PMID: 38627575]
Nicholas D Nolan, Xuan Cui, Brian M Robbings, Aykut Demirkol, Kriti Pandey, Wen-Hsuan Wu, Hannah F Hu, Laura A Jenny, Chyuan-Sheng Lin, Daniel T Hass, Jianhai Du, James B Hurley, Stephen H Tsang CRISPR editing of anti-anemia drug target rescues independent preclinical models of retinitis pigmentosa. Cell reports. Medicine 2024-04-19 [PMID: 38518771]
Jaehyeon Jeong, Byeong-Hee Kang, Sangmin Ju, Na Yeon Park, Deukyeong Kim, Ngoc Thi Bao Dinh, Jeongho Lee, Chang Yun Rhee, Dong-Hyung Cho, Hangeun Kim, Dae Kyun Chung, Heeyoun Bunch Lactiplantibacillus plantarum K8 lysates regulate hypoxia-induced gene expression Scientific Reports 2024-03-15 [PMID: 38491188]
Lassi Luomala, Kalle Mattila, Paula Vainio, Harry Nisén, Teijo Pellinen, Jouni Lohi, Teemu D. Laajala, Petrus Järvinen, Anna‐Riina Koskenniemi, Panu Jaakkola, Tuomas Mirtti Low nuclear expression of HIF ‐hydroxylases PHD2 / EGLN1 and PHD3 / EGLN3 are associated with poor recurrence‐free survival in clear cell renal cell carcinoma Cancer Medicine 2024-02-24 [PMID: 38400673]
Jenna Kerry, Erin J Specker, Morgan Mizzoni, Andrea Brumwell, Leslie Fell, Jenna Goodbrand, Michael N Rosen, James Uniacke Autophagy-dependent alternative splicing of ribosomal protein S24 produces a more stable isoform that aids in hypoxic cell survival. FEBS letters 2024-03-12 [PMID: 38281767]
Show All 822 Publications.

Reviews for HIF-2 alpha/EPAS1 Antibody (NB100-122) (39) 4.439

Average Rating: 4.4
(Based on 39 reviews)
We have 39 reviews tested in 4 species: Human, Mouse, Rat, Other.

Reviews using NB100-122:
Filter by Applications
WB
(33)
ChIP
(3)
IHC-P
(2)
IF
(1)
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Filter by Species
Human
(27)
Mouse
(6)
Rat
(1)
Other
(4)
All Species
Images Ratings Applications Species Date Details
  5
reviewed by:
Verified Customer
WB Mouse 01/11/2024
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Summary

ApplicationWestern Blot
Sample Testedmouse liver
SpeciesMouse
LotC7
Pub Med ID38652538

Comments

CommentsNuclear extracts using Nuclear and Cytoplasmic Extraction Reagents
  5
reviewed by:
Daiana Drehmer
WB Human 10/05/2022
View

Summary

ApplicationWestern Blot
Sample TestedHep3B human hepatocellular carcinoma cell line
SpeciesHuman
LotCO-5
Western Blot HIF-2 alpha/EPAS1 NB100-122
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5
reviewed by:
Verified Customer
WB Human 03/15/2021
View

Summary

ApplicationWestern Blot
Sample TestedHep3B human hepatocellular carcinoma cell line
SpeciesHuman
LotCI-3
  5
reviewed by:
Verified Customer
WB Human 07/05/2019
View

Summary

ApplicationWestern Blot
Sample TestedHuman cancer cell lines
SpeciesHuman
LotCF
Western Blot HIF-2 alpha/EPAS1 NB100-122
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5
reviewed by:
Verified Customer
WB Human 04/16/2019
View

Summary

ApplicationWestern Blot
Sample TestedHBE-16
SpeciesHuman
LotCD-9
  5
reviewed by:
Verified Customer
WB Human 02/04/2019
View

Summary

ApplicationWestern Blot
Sample Testedcultured cells
SpeciesHuman
LotBv-02
  4
reviewed by:
Verified Customer
WB Human 02/02/2019
View

Summary

ApplicationWestern Blot
Sample Tested786O cell lysates
SpeciesHuman
LotBP2
Chromatin Immunoprecipitation HIF-2 alpha/EPAS1 NB100-122
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3
reviewed by:
Verified Customer
ChIP Human 12/02/2018
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Summary

ApplicationChromatin Immunoprecipitation
Sample TestedMCF7 cell line
SpeciesHuman
LotCD-13
Western Blot HIF-2 alpha/EPAS1 NB100-122
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4
reviewed by:
Verified Customer
WB Human 09/18/2018
View

Summary

ApplicationWestern Blot
Sample TestedHuman Cancer Cells
SpeciesHuman
LotN/A
Western Blot HIF-2 alpha/EPAS1 NB100-122
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5
reviewed by:
Verified Customer
WB Human 05/11/2018
View

Summary

ApplicationWestern Blot
Sample Tested293T HEK cell lysate
SpeciesHuman
LotCD-2

Comments

CommentsAntibody dilution- 1:500 in 5% BSA- overnight 4 degerees
  5
reviewed by:
Shanhai Xie
WB Mouse 01/12/2017
View

Summary

ApplicationWestern Blot
Sample Testedtransfected U2OS cell lysate
SpeciesMouse
LotBV-2
  5
reviewed by:
Verified Customer
WB Human 01/12/2017
View

Summary

ApplicationWestern Blot
Sample Testedcultured cells
SpeciesHuman
LotBV-2
Western Blot HIF-2 alpha/EPAS1 NB100-122
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5
reviewed by:
Verified Customer
WB Mouse 10/18/2016
View

Summary

ApplicationWestern Blot
Sample TestedMouse CD4+ T Cells
SpeciesMouse
LotBP-1

Comments

CommentsDilution (1:1000)
Western Blot HIF-2 alpha/EPAS1 NB100-122
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5
reviewed by:
Helder Andre
WB Human 07/11/2016
View

Summary

ApplicationWestern Blot
Sample TestedRetinal and Choroidal primary endothelia
SpeciesHuman
Western Blot HIF-2 alpha/EPAS1 NB100-122
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5
reviewed by:
Verified Customer
WB Human 03/31/2016
View

Summary

ApplicationWestern Blot
Sample TestedHCT116 cell line lysate
SpeciesHuman
LotBP-2
Western Blot HIF-2 alpha/EPAS1 NB100-122
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4
reviewed by:
Verified Customer
WB Human 11/10/2015
View

Summary

ApplicationWestern Blot
Sample Testedcytoplasmic and nuclear protein fractions of U266 cells
SpeciesHuman
Western Blot HIF-2 alpha/EPAS1 NB100-122
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4
reviewed by:
Verified Customer
WB Human 08/12/2015
View

Summary

ApplicationWestern Blot
Sample TestedPrimary Epithelial Cells
SpeciesHuman
Western Blot HIF-2 alpha/EPAS1 NB100-122
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5
reviewed by:
Verified Customer
WB Human 06/01/2015
View

Summary

ApplicationWestern Blot
Sample TestedMDA-MB-231 Cell Lysate
SpeciesHuman
LotBQ1
  4
reviewed by:
Verified Customer
WB Human 03/27/2015
View

Summary

ApplicationWestern Blot
Sample Tested786O cell lysates
SpeciesHuman
  4
reviewed by:
Verified Customer
ChIP Human 12/12/2014
View

Summary

ApplicationChromatin Immunoprecipitation
Sample TestedSee PMID:24248342
SpeciesHuman
  4
reviewed by:
Verified Customer
WB Human 12/12/2014
View

Summary

ApplicationWestern Blot
Sample TestedSee PMID:24308012
SpeciesHuman
Western Blot HIF-2 alpha/EPAS1 NB100-122
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4
reviewed by:
Xiuquan Luo
WB Human 09/12/2014
View

Summary

ApplicationWestern Blot
Sample TestedHEK293 cell lysate
SpeciesHuman
Western Blot HIF-2 alpha/EPAS1 NB100-122
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4
reviewed by:
Pramod Mallikarjuna
WB Human 08/30/2014
View

Summary

ApplicationWestern Blot
Sample TestedClear cell renal cell carcinoma samples
SpeciesHuman
  4
reviewed by:
Verified Customer
WB Other 07/26/2014
View

Summary

ApplicationWestern Blot
Sample TestedPC12 whole cell lysate
SpeciesOther
LotBR-1
  5
reviewed by:
Verified Customer
WB Human 06/30/2014
View

Summary

ApplicationWestern Blot
Sample TestedWhole cell lysate
SpeciesHuman
Western Blot HIF-2 alpha/EPAS1 NB100-122
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5
reviewed by:
Verified Customer
WB Human 04/30/2014
View

Summary

ApplicationWestern Blot
Sample Testedwhole cell lysate U87MG (human glioblastoma cells)
SpeciesHuman
Immunohistochemistry-Paraffin HIF-2 alpha/EPAS1 NB100-122
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4
reviewed by:
Saori Yoshii
IHC-P Mouse 04/11/2014
View

Summary

ApplicationImmunohistochemistry-Paraffin
Sample TestedMouse Intestine
SpeciesMouse
LotBP-1
  4
reviewed by:
Verified Customer
WB Other 04/04/2014
View

Summary

ApplicationWestern Blot
Sample TestedSH-SY5Y whole cell lysate
SpeciesOther
LotBP-1
Western Blot HIF-2 alpha/EPAS1 NB100-122
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5
reviewed by:
Verified Customer
WB Human 04/03/2014
View

Summary

ApplicationWestern Blot
SpeciesHuman
LotBP-1
Western Blot HIF-2 alpha/EPAS1 NB100-122
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4
reviewed by:
Prabir Chakraborty
WB Human 01/24/2014
View

Summary

ApplicationWestern Blot
Sample TestedUNTREATED OVARIAN SURFACE EPITHELIUM CELLS (OSE) AND A2780 CELLS LYSATES 25ug
SpeciesHuman
LotBB1
Western Blot HIF-2 alpha/EPAS1 NB100-122
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4
reviewed by:
Verified Customer
WB Human 11/20/2013
View

Summary

ApplicationWestern Blot
Sample TestedHuman Glioma cells, whole cell lysates
SpeciesHuman
LotBN-1
Immunofluorescence HIF-2 alpha/EPAS1 NB100-122
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4
reviewed by:
Verified Customer
IF Mouse 04/15/2013
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Summary

ApplicationImmunofluorescence
SpeciesMouse
Chromatin Immunoprecipitation HIF-2 alpha/EPAS1 NB100-122
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5
reviewed by:
Verified Customer
ChIP Rat 04/02/2013
View

Summary

ApplicationChromatin Immunoprecipitation
SpeciesRat
Western Blot HIF-2 alpha/EPAS1 NB100-122
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4
reviewed by:
Verified Customer
WB 09/19/2012
View

Summary

ApplicationWestern Blot
LotBK-2
  5
reviewed by:
Verified Customer
WB Mouse 12/08/2011
View

Summary

ApplicationWestern Blot
Sample TestedMouse
SpeciesMouse
Western Blot HIF-2 alpha/EPAS1 NB100-122
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5
reviewed by:
Olga Razorenova
WB Human 06/15/2011
View

Summary

ApplicationWestern Blot
Sample TestedACHN, SN12C, Caki1 whole cell lysates, Sample Amount: 30ug
SpeciesHuman
LotBC3
  3
reviewed by:
Verified Customer
WB Other 03/11/2010
View

Summary

ApplicationWestern Blot
Sample TestedColon Cancer Cell llines, Sample Amount: 50ug
SpeciesOther
LotAT1
  3
reviewed by:
Verified Customer
WB Other 02/09/2009
View

Summary

ApplicationWestern Blot
Sample Testedhead and neck squamous cell carcinoma cells, Sample Amount: 35 ug
SpeciesOther
  3
reviewed by:
Verified Customer
IHC-P Human 02/02/2009
View

Summary

ApplicationImmunohistochemistry-Paraffin
Sample Testedhuman melanoma
SpeciesHuman
LotAI

Product General Protocols

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for HIF-2 alpha/EPAS1 Antibody (NB100-122). (Showing 1 - 10 of 10 FAQs).

  1. What is the synthetic peptide of NB100-122, corresponding to which amino acids? Can this antibody cross-react with chicken or not?
    • Unfortunately we cannot provide any further information on this epitope as it is considered proprietary. The immunogen is a peptide derived from the C-terminus of mouse/human HIF-2 alpha protein. I have carried out a blast for the % homology between human HIF-2 alpha and Chicken HIF-2 alpha. The % homology is 75% and our lab only recommends use when the homology is 85% or above.
  2. What do you recommend for blocking for a Western blot with this antibody?
    • We recommend to block membranes for 1.5 hours with 1X western wash buffer containing 5% non-fat dry milk (NFDM). For more information, a product specific WB protocol can be found on the datasheet if you click on the "Protocols & FAQs" tab.
  3. We are using HIF2 Antibody from Novus to detect HIF2 in hypoxic HUVEC cells. What washing buffer do you use for Western Blot with HIF2? Are you using PBS or TBS, 0.3 or 0.1% Tween to solute the 5% milk (NFDM)? Do you solute the antibody 1:500 or 1:1000?
    • Please see our specific protocol for Hif2 here. We recommend 0.5 to 1.0% Tween as a washing buffer and we recommend a dilution of 1:200 to 1:1000. This is depending on your sample and you will have to optimize this accordingly.
  4. I am working on a WB using the antibody against mouse HIF1a and HIF2a (catalog numbers NB100-105 and NB100-122). I tried several anti-mouse and anti-rabbit HRP secondary antibodies (from another company) and found they gave different patterns. Please provide the secondary antibody info which was used in the WB showed on the datasheet.
    • We use catalog numbers NB7570 and NB7183 as our secondary antibodies for testing these antibodies in WB.
  5. Is this antibody useful for EMSA-supershift?
    • Unfortunately we have not tested if this antibody can be used in EMSA-supershift. If you would be interested in testing this novel application, please take a look at our Innovator's Reward program.
  6. Will the hif protein be degraded when preparing the nuclear extract?
    • Working with Hif-2 can be tricky at times and the possibility of degradation is a very real one. Please see this link for a specific protocol for using NB100-122. The nuclear extract protocol can be found using this reference: Wang and Semenza. Purification and Characterization of Hypoxia-Inducible Factor 1. Journal of Biological Chemistry. 270(3): 1230-1237, 1995. I would also recommend that you use a positive control; a nuclear extract of hypoxia induced cell lines (293, Hep3B, COS7, Hepa) are all excellent choices. You can purchase the hypoxic COS7 lysate from us to make it more convenient - the catalog number is NB800-PC26. Please also see our Hypoxia support page for suggestions.
  7. We've purchased NB100-122 for Western blot and IHC. I'd like to know what would be good positive control for this antibody in case of IHC.
    • For HIF2 alpha positive control tissues, we recommend the following: Kidney, Ling, Brain, Heart. For western blot, we recommend using a positive and negative control such as normoxic/hypoxic or treated/untreated with CoCl2. We sell a CoCl2 treated/untreated Cos7 lysate: View Cos7 lysate datasheet.
  8. What is the sequence for the peptide used to purify this antibody?
    • The exact sequence of the immunogen is proprietary, but I can tell you the range is within 625-695.
  9. Could you tell us if these antibodies need to use cell lysate treated with special extraction for WB? Do you have the recommended preparation method of cell lysate?
    • Usually HIF1 alpha and HIF2 alpha are hard to detect endogenously and require some inducement. The most common is hypoxic treatment of the cells. This can be also be chemically induced by CoCl2 treatment as well. Our recommended protocols and lysate preparations can be found in our white paper here
  10. For paraffin sections, do you recommend the antigen retrieval?
    • Unless specially noted in the protocol section we always recommend using citrate buffer pH 6 for antigen retrieval. 

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Blogs on HIF-2 alpha/EPAS1.

Breast cancer stem cells survive chemotherapy through S100A10-ANXA2-SPT6 interaction that epigenetically promotes OCT4-mediated stemness
By Jamshed Arslan, Pharm D, PhDBreast cancer is the most common cancer among women that causes the greatest number of cancer-related deaths worldwide. After radiotherapy or cytotoxic chemotherapy like paclitax...  Read full blog post.

HIF-2 alpha: HIF1A's Homologue with Similar and Divergent Functions
HIF-2 alpha is a member of the heterodimeric hypoxia-inducible factors/HIFs family (HIF-1, HIF-2, and HIF-3) which contains a common beta subunit but differ in their alpha subunits. Also called as EPAS1 or Mop2, HIF-2 alpha regulates cellular adapt...  Read full blog post.

HIF-2 alpha, Tumor Suppression and Cell Survival
HIF-2 alpha is one subunit within the HIF-2 nuclear protein that regulates cellular responses to hypoxia (low oxygen tension conditions). Hydroxylation post-translational modifications on particular HIF residues target them for degradation. Luo, et al...  Read full blog post.

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Bioinformatics

Gene Symbol EPAS1