Revision 6

#48085Store 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, IHC-Bond, IHC-P, IF-IC

REACTIVITY:

H M

SENSITIVITY:

Endogenous

MW (kDa):

120

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q16665

Entrez-Gene Id:

3091

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
IHC Leica Bond 1:400 - 1:1600
Immunohistochemistry (Paraffin) 1:100 - 1:400
Immunofluorescence (Immunocytochemistry) 1:1600 - 1:6400

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.

For a carrier free (BSA and azide free) version of this product see product #87697.

Specificity / Sensitivity

HIF-1α (E1V6A) Rabbit mAb recognizes endogenous levels of total HIF-1α protein. This antibody does not cross-react with HIF-2α protein.

Species Reactivity:

Human, Mouse

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala475 of human HIF-1α protein.

Background

Hypoxia-inducible factor 1 (HIF1) is a heterodimeric transcription factor that plays a critical role in the cellular response to hypoxia (1). The HIF1 complex consists of two subunits, HIF-1α and HIF-1β, which are basic helix-loop-helix proteins of the PAS (Per, ARNT, Sim) family (2). HIF1 regulates the transcription of a broad range of genes that facilitate responses to the hypoxic environment, including genes regulating angiogenesis, erythropoiesis, cell cycle, metabolism, and apoptosis. The widely expressed HIF-1α is typically degraded rapidly in normoxic cells by the ubiquitin/proteasomal pathway. Under normoxic conditions, HIF-1α is proline hydroxylated leading to a conformational change that promotes binding to the von Hippel-Lindau protein (VHL) E3 ligase complex; ubiquitination and proteasomal degradation follows (3,4). Both hypoxic conditions and chemical hydroxylase inhibitors (such as desferrioxamine and cobalt) inhibit HIF-1α degradation and lead to its stabilization. In addition, HIF-1α can be induced in an oxygen-independent manner by various cytokines through the PI3K-AKT-mTOR pathway (5-7).

HIF-1β is also known as AhR nuclear translocator (ARNT) due to its ability to partner with the aryl hydrocarbon receptor (AhR) to form a heterodimeric transcription factor complex (8). Together with AhR, HIF-1β plays an important role in xenobiotics metabolism (8). In addition, a chromosomal translocation leading to a TEL-ARNT fusion protein is associated with acute myeloblastic leukemia (9). Studies also found that ARNT/HIF-1β expression levels decrease significantly in pancreatic islets from patients with type 2 diabetes, suggesting that HIF-1β plays an important role in pancreatic β-cell function (10).

  1. Sharp, F.R. and Bernaudin, M. (2004) Nat Rev Neurosci 5, 437-48.
  2. Wang, G.L. et al. (1995) Proc Natl Acad Sci U S A 92, 5510-4.
  3. Jaakkola, P. et al. (2001) Science 292, 468-72.
  4. Maxwell, P.H. et al. (1999) Nature 399, 271-5.
  5. Fukuda, R. et al. (2002) J Biol Chem 277, 38205-11.
  6. Jiang, B.H. et al. (2001) Cell Growth Differ 12, 363-9.
  7. Laughner, E. et al. (2001) Mol Cell Biol 21, 3995-4004.
  8. Walisser, J.A. et al. (2004) Proc Natl Acad Sci U S A 101, 16677-82.
  9. Salomon-Nguyen, F. et al. (2000) Proc Natl Acad Sci U S A 97, 6757-62.
  10. Gunton, J.E. et al. (2005) Cell 122, 337-49.

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 IHC-Bond: IHC Leica Bond IHC-P: Immunohistochemistry (Paraffin) 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.
SignalStain 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.

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 6
#48085

HIF-1α (E1V6A) Rabbit mAb

Western Blotting Image 1: HIF-1α (E1V6A) Rabbit mAb Expand Image
使用 HIF-1α (E1V6A) Rabbit mAb(上图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图),对未经处理 (-) 或经氯化钴 (CoCl2) (0.1 mM,4 小时;+)处理的 Hep G2 细胞的提取物、对未经处理 (-) 或经 CoCl2(0.1 mM,4 小时;+)处理的 Raji 细胞的提取物、对未经处理 (-) 或经 DMOG (1 mM, 6 小时;+)处理的 U-2 OS 细胞的提取物进行蛋白质印迹分析。
Western Blotting Image 2: HIF-1α (E1V6A) Rabbit mAb Expand Image
使用 HIF-1α (E1V6A) Rabbit mAb(上图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图),对未经处理 (-) 或经氯化钴(CoCl2)(0.1 mM,4 小时; +)处理的 C2C12 细胞的提取物进行蛋白质印迹分析。
Western Blotting Image 3: HIF-1α (E1V6A) Rabbit mAb Expand Image
使用 HIF-1α (E1V6A) Rabbit mAb(上图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图),对未经处理 (-) 或经 DMOG(1 mM,6 小时;+)的 NCI-H3122 细胞的提取物进行蛋白质印迹分析。
Western Blotting Image 4: HIF-1α (E1V6A) Rabbit mAb Expand Image
使用 HIF-1α (E1V6A) Rabbit mAb(上图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图),对 NCI-H28 和 MOLT-4 细胞的提取物进行蛋白质印迹分析。
Immunohistochemistry Image 1: HIF-1α (E1V6A) Rabbit mAb Expand Image
在 Leica® BOND Rx 上,使用 HIF-1α (E1V6A) Rabbit mAb 对石蜡包埋的人肾细胞癌组织进行免疫组织化学分析。
Immunohistochemistry Image 2: HIF-1α (E1V6A) Rabbit mAb Expand Image
在 Leica® BOND Rx 上,使用 HIF-1α (E1V6A) Rabbit mAb 对石蜡包埋的人胃肠道间质瘤组织进行免疫组织化学分析。
Immunohistochemistry Image 1: HIF-1α (E1V6A) Rabbit mAb Expand Image
使用 HIF-1α (E1V6A) Rabbit mAb 对石蜡包埋的人食管腺癌组织进行免疫组织化学分析。
Immunohistochemistry Image 2: HIF-1α (E1V6A) Rabbit mAb Expand Image
使用 HIF-1α (E1V6A) Rabbit mAb 对石蜡包埋的人子宫内膜样腺癌组织进行免疫组织化学分析。
Immunohistochemistry Image 3: HIF-1α (E1V6A) Rabbit mAb Expand Image
使用 HIF-1α (E1V6A) Rabbit mAb 对石蜡包埋的人导管乳腺癌组织进行免疫组织化学分析。
Immunohistochemistry Image 4: HIF-1α (E1V6A) Rabbit mAb Expand Image
使用 HIF-1α (E1V6A) Rabbit mAb 对石蜡包埋的人结肠癌组织进行免疫组织化学分析。
Immunohistochemistry Image 5: HIF-1α (E1V6A) Rabbit mAb Expand Image
使用 HIF-1α (E1V6A) Rabbit mAb 对石蜡包埋的 LL/2 同基因肿瘤组织进行免疫组织化学分析。
Immunohistochemistry Image 6: HIF-1α (E1V6A) Rabbit mAb Expand Image
使用 HIF-1α (E1V6A) Rabbit mAb 对石蜡包埋的 4T1 同基因乳腺肿瘤组织进行免疫组织化学分析。
Immunohistochemistry Image 7: HIF-1α (E1V6A) Rabbit mAb Expand Image
使用 HIF-1α (E1V6A) Rabbit mAb(上图)或 HIF-1α Rabbit mAb(下图)对石蜡包埋的人子宫内膜样腺癌组织(左图)和胃癌组织(右图)进行免疫组织化学分析。这两种抗体可检测人 HIF-1α 上独特的、不重叠表位。使用两种抗体获得的相似染色模式有助于确认染色特异性。
Immunohistochemistry Image 8: HIF-1α (E1V6A) Rabbit mAb Expand Image
与浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(右图)比较,使用 HIF-1α (E1V6A) Rabbit mAb(左图)对石蜡包埋的人正常食道组织进行免疫组织化学分析。
Immunohistochemistry Image 9: HIF-1α (E1V6A) Rabbit mAb Expand Image
使用 HIF-1α (E1V6A) Rabbit mAb 对未经处理(左图)或经 DMOG(1 mM,6 小时;右图)处理的石蜡包埋的 NCI-H3122 细胞沉淀物进行免疫组织化学分析。
Immunohistochemistry Image 10: HIF-1α (E1V6A) Rabbit mAb Expand Image
使用 HIF-1α (E1V6A) Rabbit mAb 对石蜡包埋的 NCI-H28 细胞沉淀物(左图,阳性)或 MOLT-4 细胞沉淀物(右图,阴性)进行免疫组织化学分析。
Immunohistochemistry Image 11: HIF-1α (E1V6A) Rabbit mAb Expand Image
使用 HIF-1α (E1V6A) Rabbit mAb 对未经处理(左图)或经氯化钴处理 (CoCl2)(100 μM,4 小时,右图)的 Raji 细胞沉淀物进行免疫组织化学分析。
Immunofluorescence Image 1: HIF-1α (E1V6A) Rabbit mAb Expand Image
使用 HIF-1α (E1V6A) Rabbit mAb(绿色)和 DyLight 650 Phalloidin #12956 对未经处理(左图)或经氯化钴(500 μM,24 小时;右图)处理的 Hep G2 细胞进行共聚焦免疫荧光分析。