Revision 4
#87179
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For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:
W, ChIP, ChIP-seq
Reactivity:
H
Sensitivity:
Endogenous
MW (kDa):
120
Source/Isotype:
Rabbit IgG
Product Usage Information
Application | Dilution |
---|---|
Western Blotting | 1:1000 |
Chromatin IP | 1:100 |
Chromatin IP-seq | 1:100 |
Storage
Specificity/Sensitivity
HIF-2α (E7K6E) Rabbit mAb recognizes endogenous levels of total HIF-2α protein. This antibody does not cross-react with HIF-1α protein.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu645 of human HIF-2α protein.
Background
Hypoxia-inducible factor (HIF) is essential for the cellular response to hypoxia (1,2). Under normoxia conditions, the α subunit of HIF is ubiquitinated by von Hippel-Lindau (VHL) protein and is degraded in the ubiquitin/proteasome pathway (1,2). Hypoxia inhibits degradation of the α subunit, which leads to its stabilization (1,2). HIF, in turn, regulates the transcription of a variety of genes that respond to hypoxia conditions (1,2). There are several isoforms of the HIF α subunit (2). Studies have found that HIF-1α and HIF-2α expression is increased in some human cancers (2). HIF-1α has both pro- and anti-proliferative activities, whereas HIF-2α does not possess anti-proliferative activity (2). Therefore, HIF-2α likely plays an important role in tumorigenesis (2,3).
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Western Blot Buffer
IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
Applications Key
W: Western Blotting ChIP: Chromatin IP
Cross-Reactivity Key
H: Human M: Mouse R: Rat Hm: Hamster Mk: Monkey Vir: Virus Mi: Mink C: Chicken Dm: D. melanogaster X: Xenopus Z: Zebrafish B: Bovine Dg: Dog Pg: Pig Sc: S. cerevisiae Ce: C. elegans Hr: Horse GP: Guinea Pig Rab: Rabbit G: Goat All: All Species Expected
Trademarks and Patents
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
SimpleChIP is a registered trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.
Limited Uses
Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.
Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.
Revision 4
#87179
HIF-2α (E7K6E) Rabbit mAb
Western blot analysis of Hep G2, 786-O, and A-498 cells, untreated (-) or treated with cobalt chloride (CoCl2) (0.1 mM, 4 hr; +), using HIF-2α (E7K6E) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Chromatin immunoprecipitations were performed with cross-linked chromatin from Hep 3B2.1-7 cells treated with cobalt chloride (CoCl2) (100 μM, overnight) and HIF-2α (E7K6E) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Library was prepared using DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figure shows binding across SERPINE1, a known target gene of HIF-2α (see additional figure containing ChIP-qPCR data).
Chromatin immunoprecipitations were performed with cross-linked chromatin from Hep 3B2.1-7 cells treated with cobalt chloride (CoCl2) (100 μM, overnight) and HIF-2α (E7K6E) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Library was prepared using DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figures show binding across chromosome 7 (upper), including SERPINE1 (lower), a known target gene of HIF-2α (see additional figure containing ChIP-qPCR data).
Revision 4
#87179
HIF-2α (E7K6E) Rabbit mAb
Chromatin immunoprecipitations were performed with cross-linked chromatin from Hep 3B2.1-7 cells treated with cobalt chloride (CoCl2) (100 μM, overnight) and either HIF-2α (E7K6E) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human PAI-1 Promoter Primers #33070 and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.