HIF-2 alpha/EPAS1 Antibody - BSA Free

Images

 
Biological Strategies: Western Blot: HIF-2 alpha/EPAS1 Antibody [NB100-122] - WNT11 is induced by hypoxia or hypoxic mimetics in different cell types. Immunoblot analyses of HeLa cells under normal air or hypoxia ...read more
Biological Strategies: Knockout Validated: HIF-2 alpha/EPAS1 Antibody [NB100-122] - HIF-1alpha is the predominant transcriptional regulator of WNT11 expression during hypoxia. EMSCs isolated from the indicated ...read more
Orthogonal Strategies: Dual RNAscope ISH-IHC: HIF-2 alpha/EPAS1 Antibody [NB100-122] - Formalin-fixed paraffin-embedded tissue sections of human placenta were probed for HIF-2 alpha/EPAS1 mRNA (ACD RNAScope ...read more
Immunohistochemistry: HIF-2 alpha/EPAS1 Antibody [NB100-122] - Expression of hypoxia-inducible alpha subunits in normal and diseased lung tissue. HIF-2alpha expression are more evident in fibroblasts from idiopathic ...read more
Simple Western: HIF-2 alpha/EPAS1 Antibody [NB100-122] - 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 ...read more
Genetic Strategies: Knockout Validated: HIF-2 alpha/EPAS1 Antibody [NB100-122] - Immunoblot validation of HIF2A and PHD2 KO clones using HIF2A (#NB100-122; dilution: 1/300), PHD2 (#NB100-137; dilution: 1/500). To ...read more
Biological Strategies: Western Blot: HIF-2 alpha/EPAS1 Antibody [NB100-122] - HIF-2 alpha in human retinal and choroidal primary endothelia lysates using .. Image from verified customer review.
Biological Strategies: Western Blot: HIF-2 alpha/EPAS1 Antibody [NB100-122] - HIF-2 alpha in MDA-MB-231 cell lysate (overexpression and endogenous samples) using . did not react to HIF-1 alpha overexpression. ...read more
Western Blot: HIF-2 alpha/EPAS1 Antibody [NB100-122] - Induced WNT11 expression with tumor hypoxia and WNT11 regulates tumor growth. Bevacizumab increased expression of HIF-1alpha and HIF-2alpha and WNT11. First 10 ...read more
Chromatin Immunoprecipitation: HIF-2 alpha/EPAS1 Antibody [NB100-122] - MDA-MB-231 cells were exposed to 20% or 1% O2 for 16 hours, and chromatin immunoprecipitation (ChIP) was performed with the indicated antibody ...read more
Biological Strategies: Western Blot: HIF-2 alpha/EPAS1 Antibody [NB100-122] - Lane 1: Cobalt chloride treated COS7 nuclear extracts. Lane 2: Untreated COS7 nuclear extracts.
Western Blot: HIF-2 alpha/EPAS1 Antibody [NB100-122] - 786-O cells without or with VHL overexpression. Image from verified customer review.
Biological Strategies: Western Blot: HIF-2 alpha/EPAS1 Antibody [NB100-122] - Analysis using the HRP conjugate of NB100-122. Detection of normoxic and hypoxic nuclear rat cell lysates.
Immunohistochemistry-Paraffin: HIF-2 alpha/EPAS1 Antibody [NB100-122] - HIF-2 alpha immunoreactivity in human cardiac myocytes stained with NB100-122.
Immunohistochemistry: HIF-2 alpha/EPAS1 Antibody [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 ...read more
Genetic Strategies: Knockdown Validated: HIF-2 alpha/EPAS1 Antibody [NB100-122] - Reduction of HIF-2alpha levels leads to protection in UV-triggered apoptosis, but not for apoptosis caused by glucose and serum ...read more

Product Details

Summary
Reactivity Hu, Mu, Rt, Fi, Ha, Pm, Rb, Re, ShSpecies Glossary
Applications WB, Simple Western, ChIP, ELISA, Flow, GS, IB, ICC/IF, IHC, IHC-Fr, IHC-P, In vitro, IP, PAGE, 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 1:10 - 1:500
  • Dual RNAscope ISH-IHC
  • ELISA 1:100 - 1:2000
  • Flow Cytometry
  • Gel Super Shift Assays
  • Immunoblotting
  • Immunocytochemistry/Immunofluorescence 1:100
  • Immunohistochemistry 1:100
  • Immunohistochemistry-Frozen
  • Immunohistochemistry-Paraffin 1:100
  • Immunoprecipitation 5 ug / 1 mg lysate
  • In vitro assay
  • Knockdown Validated
  • Knockout Validated
  • SDS-Page
  • Simple Western 1:50
  • Western Blot 1 - 2 ug/mL
Application Notes
Use in Gel Shift Super Assay reported in scientific literature (PMID: 15184875). Use in Immunoblotting reported in scientific literature (PMID: 28115701). 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.
Knock Out Validation was reported in scientific literature (PMID: 26861754).
Use In vitro assay reported in scientific literature (PMID: 24998849).
Knockdown validation reported in scientific literature (PMID: 31061092).
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 37 Reviews rated 4.4
using
NB100-122 in the following applications:

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

  • 55 publications
  • 1 publication
  • 6 publications
  • 3 publications
  • 2 publications
  • GS
    3 publications
  • IB
    21 publications
  • ICC
    1 publication
  • 27 publications
  • IHC
    39 publications
  • 7 publications
  • 37 publications
  • IP
    19 publications
  • KD
    1 publication
  • SW
    1 publication
  • WB
    470 publications

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)(713)

We have publications tested in 10 confirmed species: Human, Mouse, Rat, Golden Syrian hamster, Hamster, Mouse , Rabbit, Rat , Reptile, Sheep.

We have publications tested in 16 applications: ChIP, Co-IP, ELISA, FLOW, Flow, GS, IB, ICC, ICC/IF, IHC, IHC-Fr, IHC-P, IP, KD, SW, WB.


Filter By Application
ChIP
(55)
Co-IP
(1)
ELISA
(6)
FLOW
(3)
Flow
(2)
GS
(3)
IB
(21)
ICC
(1)
ICC/IF
(27)
IHC
(39)
IHC-Fr
(7)
IHC-P
(37)
IP
(19)
KD
(1)
SW
(1)
WB
(470)
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Human
(370)
Mouse
(185)
Rat
(29)
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(1)
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(2)
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(1)
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(1)
Rat
(1)
Reptile
(1)
Sheep
(1)
All Species
Showing Publications 1 - 10 of 713. Show All 713 Publications.
Publications using NB100-122 Applications Species
Tapio J, Halmetoja R, Dimova E et al. Contribution of HIF-P4H isoenzyme inhibition to metabolism indicates major beneficial effects being conveyed by HIF-P4H-2 antagonism Journal of Biological Chemistry Jul 1 2022 [PMID: 35787374] (WB, Mouse) WB Mouse
Han JS, Jeon YG, Oh M et al. Adipocyte HIF2 alpha functions as a thermostat via PKA C alpha regulation in beige adipocytes Nature communications Jun 7 2022 [PMID: 35672324] (IHC, Mouse) IHC Mouse
Salman S, Meyers D, Wicks E et al. HIF inhibitor 32-134D eradicates murine hepatocellular carcinoma in combination with anti-PD1 therapy J Clin Invest 44597 [PMID: 35499076] (WB, ChIP, Human, Mouse) WB, ChIP Human, Mouse
Xia QS, Wu F, Wu WB et al. Berberine reduces hepatic ceramide levels to improve insulin resistance in HFD-fed mice by inhibiting HIF-2 alpha Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie Apr 13 2022 [PMID: 35429745] (IHC-P, WB, ICC/IF, Human, Mouse) IHC-P, WB, ICC/IF Human, Mouse
Moyama C, Fujita M, Ando S et al. Stat5b inhibition blocks proliferation and tumorigenicity of glioblastoma stem cells derived from a de novo murine brain cancer model American journal of cancer research Mar 15 2022 [PMID: 35411230] (WB) WB
Wolf C, Smith S, van Wijk SJL Zafirlukast Induces VHL- and HIF-2alpha-Dependent Oxidative Cell Death in 786-O Clear Cell Renal Carcinoma Cells International journal of molecular sciences Mar 25 2022 [PMID: 35408930] (WB) WB
Matho C VHL-P138R and VHL-L163R Novel Variants: Mechanisms of VHL Pathogenicity Involving HIF-Dependent and HIF- Independent Actions Front. Endocrinol. 13: 854365. doi: 10.3389/fendo Jan 1 2022 [PMID: 35388293] (WB, Human) WB Human
Nelson LJ, Castro KE, Xu B et al. Synthetic lethality of cyclin-dependent kinase inhibitor Dinaciclib with VHL-deficiency allows for selective targeting of clear cell renal cell carcinoma Cell cycle (Georgetown, Tex.) Mar 4 2022 [PMID: 35240916] (WB, Human) WB Human
Armstrong N, Storey CM, Noll SE et al. SDHB knockout and succinate accumulation are insufficient for tumorigenesis but dual SDHB/NF1 loss yields SDHx-like pheochromocytomas Cell reports Mar 1 2022 [PMID: 35235785] (Mouse) Mouse
Qin y, Zhang J, Babapoor-Farrokhran S et al. PAI-1 is a vascular cell-specific HIF-2-dependent angiogenic factor that promotes retinal neovascularization in diabetic patients Science advances Mar 4 2022 [PMID: 35235351] (IHC-Fr, Human) IHC-Fr Human
Show All 713 Publications.

Reviews for HIF-2 alpha/EPAS1 Antibody (NB100-122) (37) 4.437

Average Rating: 4.4
(Based on 37 reviews)
We have 37 reviews tested in 4 species: Human, Mouse, Rat, Other.

Reviews using NB100-122:
Filter by Applications
Western Blot
(31)
Chromatin Immunoprecipitation
(3)
Immunohistochemistry-Paraffin
(2)
Immunofluorescence
(1)
All Applications
Filter by Species
Human
(26)
Mouse
(5)
Rat
(1)
Other
(4)
All Species
Images Ratings Applications Species Date Details
Western Blot HIF-2 alpha/EPAS1 NB100-122
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5
reviewed by:
Shaima Salman
Western Blot Human 03/15/2021
View

Summary

ApplicationWestern Blot
Sample TestedHep3B human hepatocellular carcinoma cell line
SpeciesHuman
LotCI-3
  5
reviewed by:
Debangshu Samanta
Western Blot Human 07/05/2019
View

Summary

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

Summary

ApplicationWestern Blot
Sample TestedHBE-16
SpeciesHuman
LotCD-9
  5
reviewed by:
John Van Haggent
Western Blot Human 02/04/2019
View

Summary

ApplicationWestern Blot
Sample Testedcultured cells
SpeciesHuman
LotBv-02
  4
reviewed by:
Steve Rey
Western Blot 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:
Jie Lan
Chromatin Immunoprecipitation Human 12/02/2018
View

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:
L P
Western Blot Human 09/18/2018
View

Summary

ApplicationWestern Blot
Sample TestedHuman Cancer Cells
SpeciesHuman
LotN/A
Western Blot HIF-2 alpha/EPAS1 NB100-122
Enlarge
5
reviewed by:
Priyanka Chaudhary
Western Blot 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:
Verified Customer
Western Blot Mouse 01/12/2017
View

Summary

ApplicationWestern Blot
Sample Testedtransfected U2OS cell lysate
SpeciesMouse
LotBV-2
  5
reviewed by:
Shanhai Xie
Western Blot Human 01/12/2017
View

Summary

ApplicationWestern Blot
Sample Testedcultured cells
SpeciesHuman
LotBV-2
Western Blot HIF-2 alpha/EPAS1 NB100-122
Enlarge
5
reviewed by:
JEEHO LEE
Western Blot 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
Enlarge
5
reviewed by:
Verified Customer
Western Blot Human 07/11/2016
View

Summary

ApplicationWestern Blot
Sample TestedRetinal and Choroidal primary endothelia
SpeciesHuman
Western Blot HIF-2 alpha/EPAS1 NB100-122
Enlarge
5
reviewed by:
Dan Cojocari
Western Blot 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:
Veena Mysore
Western Blot Human 11/10/2015
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Summary

ApplicationWestern Blot
Sample Testedcytoplasmic and nuclear protein fractions of U266 cells
SpeciesHuman
Western Blot HIF-2 alpha/EPAS1 NB100-122
Enlarge
4
reviewed by:
Benjamin Gerovac
Western Blot Human 08/12/2015
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Summary

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

Summary

ApplicationWestern Blot
Sample TestedMDA-MB-231 Cell Lysate
SpeciesHuman
LotBQ1
  4
reviewed by:
Qing Zhang
Western Blot Human 03/27/2015
View

Summary

ApplicationWestern Blot
Sample Tested786O cell lysates
SpeciesHuman
  4
reviewed by:
cynthia morimoto
Chromatin Immunoprecipitation Human 12/12/2014
View

Summary

ApplicationChromatin Immunoprecipitation
Sample TestedSee PMID:24248342
SpeciesHuman
  4
reviewed by:
cynthia morimoto
Western Blot Human 12/12/2014
View

Summary

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

Summary

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

Summary

ApplicationWestern Blot
Sample TestedClear cell renal cell carcinoma samples
SpeciesHuman
  4
reviewed by:
Ning Wang
Western Blot Other 07/26/2014
View

Summary

ApplicationWestern Blot
Sample TestedPC12 whole cell lysate
SpeciesOther
LotBR-1
  5
reviewed by:
rohini madireddy
Western Blot Human 06/30/2014
View

Summary

ApplicationWestern Blot
Sample TestedWhole cell lysate
SpeciesHuman
Western Blot HIF-2 alpha/EPAS1 NB100-122
Enlarge
5
reviewed by:
Wenwen Sun
Western Blot 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:
Verified Customer
Immunohistochemistry-Paraffin Mouse 04/11/2014
View

Summary

ApplicationImmunohistochemistry-Paraffin
Sample TestedMouse Intestine
SpeciesMouse
LotBP-1
  4
reviewed by:
Ning Wang
Western Blot 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
Enlarge
5
reviewed by:
Scott Welford
Western Blot Human 04/03/2014
View

Summary

ApplicationWestern Blot
SpeciesHuman
LotBP-1
Western Blot HIF-2 alpha/EPAS1 NB100-122
Enlarge
4
reviewed by:
Verified Customer
Western Blot 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
Enlarge
4
reviewed by:
anthony lemarie
Western Blot 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
Enlarge
4
reviewed by:
Shaohua Li
Immunofluorescence Mouse 04/15/2013
View

Summary

ApplicationImmunofluorescence
SpeciesMouse
Chromatin Immunoprecipitation HIF-2 alpha/EPAS1 NB100-122
Enlarge
5
reviewed by:
Liwei Xie
Chromatin Immunoprecipitation Rat 04/02/2013
View

Summary

ApplicationChromatin Immunoprecipitation
SpeciesRat
Western Blot HIF-2 alpha/EPAS1 NB100-122
Enlarge
4
reviewed by:
Damodara Vaddi
Western Blot 09/19/2012
View

Summary

ApplicationWestern Blot
LotBK-2
  5
reviewed by:
Li-Wen Chen
Western Blot Mouse 12/08/2011
View

Summary

ApplicationWestern Blot
Sample TestedMouse
SpeciesMouse
Western Blot HIF-2 alpha/EPAS1 NB100-122
Enlarge
5
reviewed by:
Verified Customer
Western Blot Human 06/15/2011
View

Summary

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

Summary

ApplicationWestern Blot
Sample TestedColon Cancer Cell llines, Sample Amount: 50ug
SpeciesOther
LotAT1
  3
reviewed by:
System Admin
Western Blot Other 02/09/2009
View

Summary

ApplicationWestern Blot
Sample Testedhead and neck squamous cell carcinoma cells, Sample Amount: 35 ug
SpeciesOther
  3
reviewed by:
System Admin
Immunohistochemistry-Paraffin Human 02/02/2009
View

Summary

ApplicationImmunohistochemistry-Paraffin
Sample Testedhuman melanoma
SpeciesHuman
LotAI

Product General Protocols

Video Protocols

WB Video Protocol
ChIP Video Protocol
ChIP Webinar
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: http://www.novusbio.com/whitepapers/cellular-response-hypoxia.html.
  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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Bioinformatics Tool for HIF-2 alpha/EPAS1 Antibody (NB100-122)

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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