Cell Signaling Technology

Product Pathways - Development

YAP (D8H1X) XP® Rabbit mAb #14074

Hippo   sc-101199   Yorkie  

No. Size Price
14074S 100 µl ( 10 western blots ) ¥3,580.00 现货查询 购买询价
14074T 20 µl ( 2 western blots ) ¥1,400.00 现货查询 购买询价
14074 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat,Hamster,Monkey, Endogenous 65-75 Rabbit IgG
IP 1:50
IHC-P 1:400
F 1:50
IF-IC 1:100
ChIP 1:50

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, IHC-P=Immunohistochemistry (Paraffin), F=Flow Cytometry, IF-IC=Immunofluorescence (Immunocytochemistry), ChIP=Chromatin IP,

Homology

Species predicted to react based on 100% sequence homology: Bovine, Horse,

Specificity / Sensitivity

YAP (D8H1X) XP® Rabbit mAb recognizes endogenous levels of total YAP protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the carboxy terminus of human YAP protein. The epitope corresponds to a region surrounding Pro435 of human YAP isoform 1. This sequence region is 100% conserved among all known isoforms of human YAP protein.

Western Blotting

Western Blotting

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.

IF-IC

IF-IC

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.

IHC-P (paraffin)

IHC-P (paraffin)

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

IHC-P (paraffin)

IHC-P (paraffin)

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

IP

IP

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.

Flow Cytometry

Flow Cytometry

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. 

IHC-P (paraffin)

IHC-P (paraffin)

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

IHC-P (paraffin)

IHC-P (paraffin)

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

IHC-P (paraffin)

IHC-P (paraffin)

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

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 NCI-2052 cells and either 10 µl of YAP (D8H1X) XP® Rabbit mAb, or 2 µl of 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.

Background

YAP (Yes-associated protein, YAP65) was 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 (5,6). 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. Upon phosphorylation at Ser127 by LATS1/2 kinases, YAP translocates to the cytoplasm, where it is sequestered through association with 14-3-3 proteins in an Akt-dependent manner (6-8).

  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. Basu, S. et al. (2003) Mol Cell 11, 11-23.
  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.

Application References

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Protocols

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For Research Use Only. Not For Use In Diagnostic Procedures.

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