Revision 3

#59973Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, ChIP

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

120

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q99814

Entrez-Gene Id:

2034

Product Information

Product Usage Information

For optimal ChIP results, use 5 μl of antibody and 10 μg of chromatin (approximately 4 × 10^6 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.

Application Dilution
Western Blotting 1:1000
Chromatin IP 1:200

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

HIF-2α (D6T8V) Rabbit mAb recognizes endogenous levels of total HIF-2α protein. This antibody does not cross-react with HIF-1α protein.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly688 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).

  1. Kaelin, W.G. (2005) Biochem Biophys Res Commun 338, 627-38.
  2. Toschi, A. et al. (2008) J Biol Chem 283, 34495-9.
  3. Gordan, J.D. and Simon, M.C. (2007) Curr Opin Genet Dev 17, 71-7.

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

WB: 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 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.
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 3
#59973

HIF-2α (D6T8V) Rabbit mAb

Western Blotting Image 1: HIF-2α (D6T8V) Rabbit mAb Expand Image
Western blot analysis of extracts from 786-O and A498 cells using HIF-2α (D6T8V) Rabbit mAb.
Western Blotting Image 2: HIF-2α (D6T8V) Rabbit mAb Expand Image
Western blot analysis of Hep G2 cells, untreated (-) or treated with cobalt chloride (CoCl2, 100 μM, 4 hr, +) using HIF-2α (D6T8V) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Chromatin Immunoprecipitation Image 1: HIF-2α (D6T8V) Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from Hep3B2.1-7 cells treated with cobalt chloride (100 μM) overnight and either HIF-2α (D6T8V) Rabbit mAb #59973 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, human EPO promoter primers, 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.