Revision 1
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

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

65-78

Source/Isotype:

Rabbit IgG

UniProt ID:

#P46937

Entrez-Gene Id:

10413

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50

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

Phospho-YAP (Ser109) (E5I9G) Rabbit mAb recognizes endogenous levels of YAP protein only when phosphorylated at Ser109. Based on sequence similarity, the antibody may recognize TAZ protein when phosphorylated at Ser66.

Species Reactivity:

Human, Mouse, Rat, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser109 of human YAP protein.

Background

YAP (Yes-associated protein, YAP65) was first identified based on its ability to associate with the SH3 domain of Yes. It also binds to other SH3 domain-containing proteins such as Nck, Crk, Src, and Abl (1). In addition to the SH3 binding motif, YAP contains a PDZ interaction motif, a coiled-coil domain, and WW domains (2-4). While initial studies of YAP all pointed towards a role in anchoring and targeting to specific subcellular compartments, subsequent studies showed that YAP is a transcriptional co-activator by virtue of its WW domain interacting with the PY motif (PPxY) of the transcription factor PEBP2 and other transcription factors (5). In its capacity as a transcriptional co-activator, YAP is now widely recognized as a central mediator of the Hippo Pathway, which plays a fundamental and widely conserved role in regulating tissue growth and organ size (6-8). Phosphorylation at multiple sites (e.g., Ser109, Ser127) by LATS kinases promotes YAP translocation from the nucleus to the cytoplasm, where it is sequestered through association with 14-3-3 proteins (7-9). These LATS-driven phosphorylation events serve to prime YAP for subsequent phosphorylation by CK1δ/ε in an adjacent phosphodegron, triggering proteasomal degradation of YAP (10).

  1. Sudol, M. (1994) Oncogene 9, 2145-52.
  2. Mohler, P.J. et al. (1999) J Cell Biol 147, 879-90.
  3. Espanel, X. and Sudol, M. (2001) J Biol Chem 276, 14514-23.
  4. Sudol, M. et al. (1995) FEBS Lett 369, 67-71.
  5. Yagi, R. et al. (1999) EMBO J 18, 2551-62.
  6. Dong, J. et al. (2007) Cell 130, 1120-33.
  7. Zhao, B. et al. (2010) Genes Dev 24, 862-74.
  8. Zhao, B. et al. (2007) Genes Dev 21, 2747-61.
  9. Yu, F.X. et al. (2012) Cell 150, 780-91.
  10. Zhao, B. et al. (2010) Genes Dev 24, 72-85.

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 IP: Immunoprecipitation

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 1
#53749

Phospho-YAP (Ser109) (E5I9G) Rabbit mAb

Western Blotting Image 1: Phospho-YAP (Ser109) (E5I9G) Rabbit mAb Expand Image
使用 Phospho-YAP (Ser109) (E5I9G) Rabbit mAb #53749(上图)和 GAPDH (D6H11) XP® Rabbit mAb #5174(下图)对 HeLa 细胞(泳道 1)或 YAP 敲除型细胞(泳道 2)的提取物进行蛋白印迹分析。YAP 敲除型 HeLa 细胞中没有信号,这证实了抗体对 YAP 的特异性。
Western Blotting Image 2: Phospho-YAP (Ser109) (E5I9G) Rabbit mAb Expand Image
使用 Phospho-YAP (Ser109) (E5I9G) 兔单克隆抗体(上图)、YAP (D8H1X) XP® 兔单克隆抗体 #14074(中间)和 β-肌动蛋白 (D6A8) 兔单克隆抗体 #8457(下图)对未经 (-) 或已经牛小肠磷酸酶 (CIP) 和 λ-磷酸酶 (+) 处理的 PANC-1 细胞和 HeLa 细胞的提取物进行蛋白印迹分析。
Western Blotting Image 3: Phospho-YAP (Ser109) (E5I9G) Rabbit mAb Expand Image
使用 Phospho-YAP (Ser109) (E5I9G) 兔单克隆抗体(上图)和 β-肌动蛋白 (D6A8) 兔单克隆抗体 #8457(下图)对不同细胞系的提取物进行蛋白印迹分析。RL 细胞的 YAP 表达呈阴性,从而证实了该抗体的特异性。正如已发表的基因表达数据库中预测以及使用其他 YAP 抗体所确认的,在 RL 提取物中无法检测到 YAP 表达。
Immunoprecipitation Image 1: Phospho-YAP (Ser109) (E5I9G) Rabbit mAb Expand Image
对 PANC-1 细胞提取物磷酸化 YAP (Ser109) 蛋白进行免疫沉淀。泳道 1 为 10% 敲入,泳道 2 为兔 (DA1E) 单克隆抗体 IgG XP® 同型对照 #3900,泳道 3 为 Phospho-YAP (Ser109) (E5I9G) 兔单克隆抗体。使用 Phospho-YAP (Ser109) 抗体 #46931 进行蛋白印迹分析。小鼠抗兔 IgG (构象特异) (L27A9) 单克隆抗体 (HRP 偶联物) #5127 用于检测,以免与 IgG 发生交叉反应。