WB, IP
H M R Mk
Endogenous
78
Rabbit
#P46937
10413
Product Information
Product Usage Information
| Application | Dilution |
|---|---|
| Western Blotting | 1:1000 |
| Immunoprecipitation | 1:200 |
Storage
Specificity / Sensitivity
Species Reactivity:
Human, Mouse, Rat, Monkey
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser61 of human YAP protein. Antibodies are purified by peptide affinity chromatography.
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).
- Sudol, M. (1994) Oncogene 9, 2145-52.
- Mohler, P.J. et al. (1999) J Cell Biol 147, 879-90.
- Espanel, X. and Sudol, M. (2001) J Biol Chem 276, 14514-23.
- Sudol, M. et al. (1995) FEBS Lett 369, 67-71.
- Yagi, R. et al. (1999) EMBO J 18, 2551-62.
- Dong, J. et al. (2007) Cell 130, 1120-33.
- Zhao, B. et al. (2010) Genes Dev 24, 862-74.
- Zhao, B. et al. (2007) Genes Dev 21, 2747-61.
- Yu, F.X. et al. (2012) Cell 150, 780-91.
- Zhao, B. et al. (2010) Genes Dev 24, 72-85.
- Mo, J.S. et al. (2015) Nat Cell Biol 17, 500-10.
- Wang, W. et al. (2015) Nat Cell Biol 17, 490-9.
- 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 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
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