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52420
Phospho-YAP/TAZ Antibody Sampler Kit
Primary Antibodies

Phospho-YAP/TAZ Antibody Sampler Kit #52420

Western Blotting Image 1

Western blot analysis of extracts from PANC-1 cells (human) and NIH/3T3 cells (mouse), untreated (-) or treated with λ phosphatase (+), using Phospho-YAP (Ser109) Antibody (upper) and YAP (D8H1X) XP® Rabbit mAb #14074 (lower).

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Western Blotting Image 2

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).

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Western Blotting Image 3

Western blot analysis of extracts from Hep G2 cells, untreated (-) or λ-phosphatase-treated (+), using Phospho-YAP (Ser397) (D1E7Y) Rabbit mAb (upper), YAP Antibody #4912 (middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower). YAP protein isoform 1 Ser397 corresponds to Ser381 of YAP isoform 2, as reported by Zhao et al. (2010) Genes Dev 24, 72-85 (9).

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Western Blotting Image 4

Western blot analysis of extracts from PANC-1 and A-204 cells, untreated (-) or treated with calf intestinal phosphatase (CIP) and λ phosphatase (+) using Phospho-TAZ (Ser89) (E1X9C) Rabbit mAb (upper), TAZ (D3I6D) Rabbit MAb #70148 (middle) and β-Actin (D6A8) Rabbit mAb #8457 (lower). Asterisk (*) indicates weak detection of phosphorylated YAP, due to sequence similarity in the regions surrounding TAZ (Ser89) and YAP (Ser127).

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Western Blotting Image 5

Western blot analysis of extracts from various cell lines using YAP/TAZ (D24E4) Rabbit mAb.

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Western Blotting Image 6

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.

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Chromatin IP-seq Image 7

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). For additional ChIP-seq tracks, please download the product data sheet.

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Western Blotting Image 8

Western blot analysis of extracts from various cells using TAZ (D3I6D) Rabbit mAb (upper) and GAPDH (D16H11) XP® #5174 (lower). As expected, Raji cells are negative for TAZ.

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Chromatin IP-seq Image 9

Chromatin immunoprecipitations were performed with cross-linked chromatin from NCI-H2052 cells and TAZ (D3I6D) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across SMYD3, a known target gene of TAZ (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.

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Western Blotting Image 10

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

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Western Blotting Image 11

Western blot analysis of extracts from various cell lines using using Phospho-YAP (Ser109) Antibody (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). RL-7 cells are negative for YAP expression, confirming specificity of the antibody.

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Western Blotting Image 12

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].

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Western Blotting Image 13

Western blot analysis of extracts from MDA-MB-231 cells, vehicle treated (-) or treated with epinephrine (10 μM, 60 min; +) or Forskolin #3828 (10 μm, 60 min; +), using Phospho-YAP (Ser397) (D1E7Y) Rabbit mAb (upper), YAP Antibody #4912 (middle), and β-Actin (D6A8) Rabbit mAb #8547 (lower). YAP protein isoform 1 Ser397 corresponds to Ser381 of YAP isoform 2, as reported by Zhao, B. et al. (2010) Genes Dev 24, 72-85 (9).

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IP Image 14

Immunoprecipitation of Phospho-TAZ (Ser89) from A-204 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Phospho-TAZ (Ser89) (E1X9C) Rabbit mAb. Western blot analysis was performed using Phospho-TAZ (Ser89) (E1X9C) Rabbit mAb. Mouse Anti-Rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as the secondary antibody. Asterisk (*) indicates weak detection of phosphorylated YAP, due to sequence similarity in the regions surrouding TAZ (Ser89) and YAP (Ser127).

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IHC-P (paraffin) Image 15

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using YAP/TAZ (D24E4) Rabbit mAb.

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IP Image 16

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.

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Chromatin IP Image 17

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.

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Chromatin IP Image 18

Chromatin immunoprecipitations were performed with cross-linked chromatin from NCI-H2052 cells and either 10 μl of TAZ (D3I6D) Rabbit mAb or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. 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.

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IP Image 19

Immunoprecipitation of Phospho-YAP (Ser109) from PANC-1 cell extracts. Lane 1 is 10% input, lane 2 is Normal Rabbit IgG #2729, and lane 3 is Phospho-YAP (Ser109) Antibody. Western blot analysis was performed using Phospho-YAP (Ser109) Antibody.

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Western Blotting Image 20

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

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Western Blotting Image 21

Western blot analysis of extracts from various cell lines using Phospho-YAP (Ser397) (D1E7Y) Rabbit mAb.

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IHC-P (paraffin) Image 22

Immunohistochemical analysis of paraffin-embedded human lymphoma using YAP/TAZ (D24E4) Rabbit mAb.

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IHC-P (paraffin) Image 23

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

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IHC-P (paraffin) Image 24

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

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Isoform Specificity Image 25

Schematic diagram showing annotation of the amino acid sequence surrounding Ser397 in different human YAP isoforms. Asterisk (*) indicates annotation of the phosphorylation site as described in Zhao et al. 2010 [9].

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IHC-P (paraffin) Image 26

Immunohistochemical analysis of paraffin-embedded COS-7 cell pellets, transfected with YAP (left) or TAZ (right), using YAP/TAZ (D24E4) Rabbit mAb.

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IHC-P (paraffin) Image 27

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

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IHC-P (paraffin) Image 28

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

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IHC-P (paraffin) Image 29

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

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IHC-P (paraffin) Image 30

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).

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IHC-P (paraffin) Image 31

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

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IHC-P (paraffin) Image 32

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

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Flow Cytometry Image 33

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. 

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IF-IC Image 34

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.

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Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Phospho-YAP (Ser109) Antibody 46931 20 µl
  • WB
  • IP
H M R Mk 78 Rabbit 
Phospho-YAP (Ser127) (D9W2I) Rabbit mAb 13008 20 µl
  • WB
  • IP
  • IHC
H M R 65-75 Rabbit IgG
Phospho-YAP (Ser397) (D1E7Y) Rabbit mAb 13619 20 µl
  • WB
  • IP
H M R 75 Rabbit IgG
Phospho-TAZ (Ser89) (E1X9C) Rabbit mAb 59971 20 µl
  • WB
  • IP
H M R 55 Rabbit IgG
YAP/TAZ (D24E4) Rabbit mAb 8418 20 µl
  • WB
  • IP
  • IHC
H M Mk 50, 70 Rabbit IgG
YAP (D8H1X) XP® Rabbit mAb 14074 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
  • ChIP
H M R Hm Mk 65-75 Rabbit IgG
TAZ (D3I6D) Rabbit mAb 70148 20 µl
  • WB
  • IP
  • ChIP
H M 50 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

The Phospho-YAP/TAZ Antibody Sampler Kit uses phospho-specific and control antibodies to provide an economical means of detecting the phosphorylation of YAP and TAZ proteins at critical residues that are reported to regulate YAP and TAZ protein stability. The kit includes enough antibody to perform two western blot experiments with each primary antibody.

Each antibody in the Phospho-YAP/TAZ Antibody Sampler Kit detects endogenous levels of its target protein. Phospho-YAP (Ser127) (D9W2I) Rabbit mAb recognizes endogenous levels of YAP protein only when phosphorylated at Ser127. Phospho-YAP (Ser397) (D1E7Y) Rabbit mAb recognizes endogenous levels of YAP protein only when phosphorylated at Ser397. Phospho-YAP (Ser109) Antibody recognizes endogenous levels of YAP protein only when phosphorylated at Ser109. Phospho-TAZ (Ser89) (E1X9C) Rabbit mAb recognizes endogenous levels of TAZ protein only when phosphorylated at Ser89. Due to epitope sequence similarities, Phospho-TAZ (Ser89) (E1X9C) Rabbit mAb may weakly detect YAP protein, but only when YAP is phosphorylated at Ser127.

Monoclonal antibodies are produced by immunizing rabbits with recombinant protein corresponding to the carboxy terminus of human YAP protein and synthetic peptides corresponding to Ala200 of mouse TAZ protein and Asp362 of human TAZ protein. Phosphorylation-specific antibodies are produced by immunizing rabbits with synthetic phospho-peptides corresponding to Ser109, Ser127, and Ser397 of human YAP protein, and Ser89 of human TAZ protein. Polyclonal antibodies are purified by Protein A and peptide affinity chromatography.

YAP and TAZ (WWTR1) are transcriptional co-activators that play a central role in the Hippo Signaling pathway that regulates cell, tissue and organ growth. YAP and TAZ are structurally and functionally similar, but exhibit differential patterns of expression among cells and tissues that suggest partially non-redundant functions (1). YAP and TAZ are dynamically regulated in response to internal and external cellular signals. Under growth conditions, YAP and TAZ are translocated to the nucleus, where they interact with transcription factors (e.g., TEA domain proteins) that regulate the transcription of genes that control proliferation and cell survival (2). The subcellular localization of YAP and TAZ is dynamically regulated by a kinase cascade that regulates the phosphorylation status of key residues within YAP and TAZ. Phosphorylation of YAP and TAZ (e.g., Ser109, Ser127, Ser397 in YAP; Ser89 in TAZ) results in their cytoplasmic translocation, sequestration by 14-3-3 proteins, and recruitment of the β-TrCP (SCF) ubiquitin ligase complex (3,4). This complex ubiquitinates YAP and TAZ, triggering their proteolytic degradation in the proteasome, thereby altering the transcription of genes that control proliferation and cell survival (3-5).

  1. Piccolo, S. et al. (2014) Physiol Rev 94, 1287-312.
  2. Holden, J.K. and Cunningham, C.N. (2018) Cancers (Basel) 10, .
  3. Lei, Q.Y. et al. (2008) Mol Cell Biol 28, 2426-36.
  4. Zhao, B. et al. (2010) Genes Dev 24, 72-85.
  5. Cordenonsi M et al. (2011) Cell 147, 759–72
Entrez-Gene Id
25937 , 10413
Swiss-Prot Acc.
Q9GZV5 , P46937
For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.

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