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46538
PhosphoPlus® YAP (Ser127) Antibody Duet
Primary Antibodies
Antibody Duet

PhosphoPlus® YAP (Ser127) Antibody Duet #46538

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other Image 1 - PhosphoPlus® YAP (Ser127) Antibody Duet

Western blot analysis of extracts from control HeLa cells (Lane 1) or YAP knockout HeLa cells (Lane 2) using YAP (D8H1X) XP® Rabbit mAb (upper) or β-actin (13E5) Rabbit mAb #4970 (lower). The absence of signal in the YAP knockout HeLa cells confirms the specificity of the antibody for YAP.

other Image 2 - PhosphoPlus® YAP (Ser127) Antibody Duet

Chromatin immunoprecipitations were performed with cross-linked chromatin from NCI-H2052 cells and YAP (D8H1X) XP® Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across CTGF, a known target gene of YAP (see additional figure containing ChIP-qPCR data).

other Image 3 - PhosphoPlus® YAP (Ser127) Antibody Duet

Western blot analysis of extracts from HeLa cells (lane 1) or YAP knock-out cells (lane 2) using Phospho-YAP (Ser127) (D9W2I) Rabbit mAb #13008 (upper), and GAPDH (D6H11) XP® Rabbit mAb #5174 (lower). The absence of signal in the YAP knock-out HeLa cells confirms specificity of the antibody for YAP.

other Image 4 - PhosphoPlus® YAP (Ser127) Antibody Duet

Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma, control (left) or λ-phosphatase treated (right), using Phospho-YAP (Ser127) (D9W2I) Rabbit mAb.

other Image 5 - PhosphoPlus® YAP (Ser127) Antibody Duet

Western blot analysis of extracts from various cell lines using YAP (D8H1X) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). As expected, RL-7 cells are negative for YAP protein expression.

other Image 6 - PhosphoPlus® YAP (Ser127) Antibody Duet

Chromatin immunoprecipitations were performed with cross-linked chromatin from NCI-H2052 cells and YAP (D8H1X) XP® Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across chromosome 6 (upper), including CTGF (lower), a known target gene of YAP (see additional figure containing ChIP-qPCR data).

other Image 7 - PhosphoPlus® YAP (Ser127) Antibody Duet

Immunohistochemical analysis of paraffin-embedded cell pellets, A-204 (left) and RL-7 (right), using Phospho-YAP (Ser127) (D9W2I) Rabbit mAb.

other Image 8 - PhosphoPlus® YAP (Ser127) Antibody Duet

Chromatin immunoprecipitations were performed with cross-linked chromatin from NCI-2052 cells and either YAP (D8H1X) XP® Rabbit mAb, or Normal Rabbit IgG #2729, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human CTGF Promoter Primers #14927, human SMYD3 intron 2 primers, and SimpleChIP® Human CTGF Upstream Primers #14928. 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.

other Image 9 - PhosphoPlus® YAP (Ser127) Antibody Duet

Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma using Phospho-YAP (Ser127) (D9W2I) Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).

other Image 10 - PhosphoPlus® YAP (Ser127) Antibody Duet

Flow cytometric analysis of RL-7 cells (blue) and A-204 cells (green) using YAP (D8H1X) XP® Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody. 

other Image 11 - PhosphoPlus® YAP (Ser127) Antibody Duet

Confocal immunofluorescent analysis of low confluence MCF 10A cells (left), high confluence MCF 10A (center), and YAP negative RL-7 cells (right) using YAP (D8H1X) XP® Rabbit mAb (green). Blue pseudocolor in lower images = DRAQ5® #4084 (fluorescent DNA dye). Increased nuclear localization of YAP protein is seen in low confluence (proliferating) cells.

other Image 12 - PhosphoPlus® YAP (Ser127) Antibody Duet

Western blot analysis of extracts from PANC-1 cells, untreated (-) or λ-phosphatase-treated (+), using Phospho-YAP (Ser127) (D9W2I) Rabbit mAb (upper), YAP Antibody #4912 (middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower).

other Image 13 - PhosphoPlus® YAP (Ser127) Antibody Duet

Immunohistochemical analysis of paraffin-embedded human non-Hodgkin’s lymphoma using YAP (D8H1X) XP® Rabbit mAb performed on the Leica® BOND™ Rx. 

other Image 14 - PhosphoPlus® YAP (Ser127) Antibody Duet

Western blot analysis of MDA-MB-231 cells, vehicle-treated (-) or treated with Forskolin #3828 (10 μM, 60 min; +) or epinephrine (10 μM, 60 min; +), using Phospho-YAP (Ser127) (D9W2I) Rabbit mAb (upper), YAP Antibody #4912 (middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower). Note the induction of YAP (Ser127) phosphorylation after treatment with forskolin or epinephrine, consistent with the findings reported in Xu et al. (2012) [9].

other Image 15 - PhosphoPlus® YAP (Ser127) Antibody Duet

Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using YAP (D8H1X) XP® Rabbit mAb performed on the Leica® BOND™ Rx.  

other Image 16 - PhosphoPlus® YAP (Ser127) Antibody Duet

Western blot analysis of extracts from various cell lines using Phospho-YAP (Ser127) (D9W2I) Rabbit mAb.

other Image 17 - PhosphoPlus® YAP (Ser127) Antibody Duet

Immunohistochemical analysis of paraffin-embedded human breast adenocarcinoma using YAP (D8H1X) XP® Rabbit mAb.

other Image 18 - PhosphoPlus® YAP (Ser127) Antibody Duet

Immunohistochemical analysis of paraffin-embedded human breast adenocarcinoma using YAP (D8H1X) XP® Rabbit mAb.

other Image 19 - PhosphoPlus® YAP (Ser127) Antibody Duet

Immunohistochemical analysis of paraffin-embedded 293T cell pellet, wild-type (left, positive) or YAP/TAZ knockout (right, negative), using YAP (D8H1X) XP® Rabbit mAb.

other Image 20 - PhosphoPlus® YAP (Ser127) Antibody Duet

Immunohistochemical analysis of paraffin-embedded BEN cell pellet, untransfected (left, YAP/TAZ low), YAP-transfected (middle) or TAZ-transfected (right), using YAP (D8H1X) XP® Rabbit mAb (top) or TAZ (E9J5A) XP® Rabbit mAb #72804 (bottom).

other Image 21 - PhosphoPlus® YAP (Ser127) Antibody Duet

Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma (left), colon carcinoma case 1 (middle) or colon carcinoma case 2 (right), using YAP (D8H1X) XP® Rabbit mAb (top) or TAZ (E9J5A) XP® Rabbit mAb #72804 (bottom).

other Image 22 - PhosphoPlus® YAP (Ser127) Antibody Duet

Immunohistochemical analysis of paraffin-embedded human ovarian serous carcinoma using YAP (D8H1X) XP® Rabbit mAb.

other Image 23 - PhosphoPlus® YAP (Ser127) Antibody Duet

Immunohistochemical analysis of paraffin-embedded PANC-1 (left) and RL-7 (right) cell pellets using YAP (D8H1X) XP® Rabbit mAb.

other Image 24 - PhosphoPlus® YAP (Ser127) Antibody Duet

Immunohistochemical analysis of paraffin-embedded human benign prostatic hyperplasia using YAP (D8H1X) XP® Rabbit mAb.

other Image 25 - PhosphoPlus® YAP (Ser127) Antibody Duet

Immunoprecipitation of YAP protein from A-204 cell extracts using Rabbit (DA1E) mAb XP® Isotype Control #3900 (lane 2) or YAP (D8H1X) XP® Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using YAP (D8H1X) XP® Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb #3678 was used as a secondary antibody.

To Purchase # 46538S
製品番号 サイズ 価格 在庫
46538S
1 Kit

Product Includes Quantity Reactivity MW(kDa) Isotype
YAP (D8H1X) XP® Rabbit mAb 14074 100 µl H M R Hm Mk 65-78 Rabbit IgG
Phospho-YAP (Ser127) (D9W2I) Rabbit mAb 13008 100 µl H M R 65-78 Rabbit IgG

Product Description

PhosphoPlus® Duets from Cell Signaling Technology (CST) provide a means to assess protein activation status. Each Duet contains an activation-state and total protein antibody to your target of interest. These antibodies have been selected from CST's product offering based upon superior performance in specified applications.

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.

Pathways & Proteins

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