Revision 3

#71565Store 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, IP, IF-IC

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

120

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q99814

Entrez-Gene Id:

2034

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Immunofluorescence (Immunocytochemistry) 1:100 - 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 (E8E5Z) 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 Glu600 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 BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IF-IC: Immunofluorescence (Immunocytochemistry)

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.
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.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品, (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Revision 3
#71565

HIF-2α (E8E5Z) Rabbit mAb

Western Blotting Image 1: HIF-2α (E8E5Z) Rabbit mAb Expand Image
使用 HIF-2α (E8E5Z) Rabbit mAb(上图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图)对经过赋形剂处理 (-) 或经过氯化钴(CoCl2,500 μM,18 小时; +)处理的 Hep G2 细胞进行蛋白质印迹分析。
Western Blotting Image 2: HIF-2α (E8E5Z) Rabbit mAb Expand Image
使用 HIF-2α (E8E5Z) Rabbit mAb(上图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图)对经过赋形剂处理或经过氯化钴(CoCl2,0.1 mM,4 小时;+)处理的 Hep G2 细胞、未经处理的 786-O 和 A-498 细胞进行蛋白质印迹分析。
Immunoprecipitation Image 1: HIF-2α (E8E5Z) Rabbit mAb Expand Image
对 786-O 细胞提取物中的 HIF-2α 蛋白进行免疫沉淀分析。泳道 1 为 10% input,泳道 2 为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,泳道 3 为 HIF-2α (E8E5Z) Rabbit mAb。使用 HIF-2α (E8E5Z) Rabbit mAb 进行蛋白质印迹分析。Anti-rabbit IgG, HRP-linked Antibody #7074 用作二抗。
Immunofluorescence Image 1: HIF-2α (E8E5Z) Rabbit mAb Expand Image
使用 HIF-2α (E8E5Z) Rabbit mAb(绿色)、DyLight 554 Phalloidin #13054(红色)和 DAPI #4083(蓝色)对经过赋形剂处理(左图)或经过氯化钴(500 μM,18 小时;右图)处理的 Hep G2 细胞进行共聚焦免疫荧光分析。