Revision 3
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, W-S, IP, IHC-P

REACTIVITY:

H M R

SENSITIVITY:

Endogenous

MW (kDa):

78

Source/Isotype:

Rabbit IgG

UniProt ID:

#P46937

Entrez-Gene Id:

10413

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:10 - 1:50
Immunoprecipitation 1:100
Immunohistochemistry (Paraffin) 1:1000 - 1:4000

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 (Ser61) (E4Q8S) Rabbit mAb recognizes endogenous levels of YAP protein only when phosphorylated at Ser61.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser61 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).
YAP function is influenced by cellular metabolic status. This was recognized after observing that AMPK, a master regulator of cell metabolism, directly phosphorylates YAP at multiple sites (e.g., Ser61, Ser94) during energy stress or nutritional deprivation (11-13). Phosphorylation of Ser61 by AMPK did not increase 14-3-3 binding or affect YAP subcellular localization, but nevertheless resulted in transcriptional repression of YAP target genes (12).

  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.
  11. Mo, J.S. et al. (2015) Nat Cell Biol 17, 500-10.
  12. Wang, W. et al. (2015) Nat Cell Biol 17, 490-9.
  13. Hariharan, I.K. (2015) Nat Cell Biol 17, 362-3.

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

WB: Western Blotting W-S: Simple Western™ IP: Immunoprecipitation IHC-P: Immunohistochemistry (Paraffin)

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.
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
#86312

Phospho-YAP (Ser61) (E4Q8S) Rabbit mAb

Western Blotting Image 1: Phospho-YAP (Ser61) (E4Q8S) Rabbit mAb Expand Image
使用 Phospho-YAP (Ser61) (E4Q8S) Rabbit mAb(上图)、YAP (D8H1X) XP® Rabbit mAb #14074(中图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图)对 293T 细胞(泳道 1)或 293T/YAP/TAZ 敲除型 (KO) 细胞(泳道 2)的提取物进行蛋白质印迹分析。KO 细胞的提取物中没有信号证实了抗体对 YAP 的特异性。YAP/TAZ KO 细胞由波士顿儿童医院 Fernando Camargo 博士惠赠。
Western Blotting Image 2: Phospho-YAP (Ser61) (E4Q8S) Rabbit mAb Expand Image
使用 Phospho-YAP (Ser61) (E4Q8S) Rabbit mAb(上图)、YAP (D8H1X) XP® Rabbit mAb #14074(中图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图)对未经处理 (-) 或已经过 λ-磷酸酶/牛小肠磷酸酶 (CIP) (+) 处理的 PANC-1 细胞的提取物进行蛋白质印迹分析。
Western Blotting Image 1: Phospho-YAP (Ser61) (E4Q8S) Rabbit mAb Expand Image
Simple Western™ analysis of lysates (1.0 mg/mL) from HeLa cells using Phospho-YAP (Ser61) (E4Q8S) Rabbit mAb #86312. 虚拟泳道式图像(左图)显示一抗稀释比例为 1:10 和 1:50 时的靶标条带(如图所示)。对应的电泳图(右图)为一抗稀释比例在 1:10(蓝线)和 1:50(绿线)时沿毛细血管内分子量的化学发光结果。在还原条件下,使用 12-230 kDa 分离模块在 ProteinSimple(BioTechne 品牌)的 Jess™ Simple Western 仪器上进行该实验。
Immunoprecipitation Image 1: Phospho-YAP (Ser61) (E4Q8S) Rabbit mAb Expand Image
对 PANC-1 细胞提取物中的磷酰-YAP (Ser61) 蛋白进行免疫沉淀分析。泳道 1 为 10% 输入,泳道 2 为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,泳道 3 为 Phospho-YAP (Ser61) (E4Q8S) Rabbit mAb。使用 Phospho-YAP (Ser61) Antibody #75784 进行蛋白质印迹分析。Anti-rabbit IgG, HRP-linked Antibody #7074 用作二抗。
Immunohistochemistry Image 1: Phospho-YAP (Ser61) (E4Q8S) Rabbit mAb Expand Image
使用 Phospho-YAP (Ser61) (E4Q8S) 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: Phospho-YAP (Ser61) (E4Q8S) Rabbit mAb Expand Image
使用 Phospho-YAP (Ser61) (E4Q8S) Rabbit mAb 对石蜡包埋的人鳞状细胞肺癌组织进行免疫组织化学分析。
Immunohistochemistry Image 3: Phospho-YAP (Ser61) (E4Q8S) Rabbit mAb Expand Image
使用 Phospho-YAP (Ser61) (E4Q8S) Rabbit mAb 对石蜡包埋的人前列腺癌组织进行免疫组织化学分析。
Immunohistochemistry Image 4: Phospho-YAP (Ser61) (E4Q8S) Rabbit mAb Expand Image
使用 Phospho-YAP (Ser61) (E4Q8S) Rabbit mAb 对石蜡包埋的人子宫内膜腺癌组织进行免疫组织化学分析。
Immunohistochemistry Image 5: Phospho-YAP (Ser61) (E4Q8S) Rabbit mAb Expand Image
使用 Phospho-YAP (Ser61) (E4Q8S) 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 6: Phospho-YAP (Ser61) (E4Q8S) Rabbit mAb Expand Image
使用 Phospho-YAP (Ser61) (E4Q8S) Rabbit mAb 对石蜡包埋的人胸腺癌肉瘤组织进行免疫组织化学分析。
Immunohistochemistry Image 7: Phospho-YAP (Ser61) (E4Q8S) Rabbit mAb Expand Image
使用 Phospho-YAP (Ser61) (E4Q8S) Rabbit mAb 对石蜡包埋的人正常小肠组织进行免疫组织化学分析。
Immunohistochemistry Image 8: Phospho-YAP (Ser61) (E4Q8S) Rabbit mAb Expand Image
使用 Phospho-YAP (Ser61) (E4Q8S) Rabbit mAb 对石蜡包埋的人正常胎盘组织进行免疫组织化学分析。
Immunohistochemistry Image 9: Phospho-YAP (Ser61) (E4Q8S) Rabbit mAb Expand Image
使用 Phospho-YAP (Ser61) (E4Q8S) Rabbit mAb 对石蜡包埋的 Renca 同基因肿瘤组织进行免疫组织化学分析。
Immunohistochemistry Image 10: Phospho-YAP (Ser61) (E4Q8S) Rabbit mAb Expand Image
使用 Phospho-YAP (Ser61) (E4Q8S) 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 11: Phospho-YAP (Ser61) (E4Q8S) Rabbit mAb Expand Image
使用 Phospho-YAP (Ser61) (E4Q8S) Rabbit mAb 对石蜡包埋的 A20 同基因肿瘤组织进行免疫组织化学分析。
Immunohistochemistry Image 12: Phospho-YAP (Ser61) (E4Q8S) Rabbit mAb Expand Image
使用 Phospho-YAP (Ser61) (E4Q8S) Rabbit mAb 对石蜡包埋的小鼠脾组织进行免疫组织化学分析。
Immunohistochemistry Image 13: Phospho-YAP (Ser61) (E4Q8S) Rabbit mAb Expand Image
使用 Phospho-YAP (Ser61) (E4Q8S) Rabbit mAb 对未经处理(左图)或经 λ 磷酸酶处理(右图)的石蜡包埋的人正常胰腺组织进行免疫组织化学分析。
Immunohistochemistry Image 14: Phospho-YAP (Ser61) (E4Q8S) Rabbit mAb Expand Image
使用 Phospho-YAP (Ser61) (E4Q8S) Rabbit mAb 对石蜡包埋的野生型 293T 细胞沉淀物(左图,阳性)或 YAP/TAZ 敲除型 (KO) 293T 细胞沉淀物(右图,阴性)进行免疫组织化学分析。
Immunohistochemistry Image 15: Phospho-YAP (Ser61) (E4Q8S) Rabbit mAb Expand Image
使用 Phospho-YAP (Ser61) (E4Q8S) Rabbit mAb 对石蜡包埋的未转染(左图)、YAP 转染(中图)或 TAZ 转染(右图)的 BEN 细胞沉淀物进行免疫组织化学分析。