YAP
| Cat Number: | MAB-94469 |
|---|---|
| Size: | 100μg |
| Clone: | D8H1X |
| Concentration: | 1mg/ml |
| Host: | Rb |
| Isotype: | IgG |
| Reactivity: | Hu,Ms,Rt |
| Applications: | Western blotting 1:1000 |
| Molecular Weight: | 65-75 kDa |
| 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. |
| 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 coactivator 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 coactivator, 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).YAP (D8H1X) XP® Rabbit mAb recognizes endogenous levels of total YAP protein. |
| Form: | Liquid |
| Buffer: | PBS with 0.02%sodium azide, 50% glycerol, pH7.3. |
Western blot analysis of extracts of
various cell lines, using YAP1 antibody
at 1:1000 dilution. Secondary antibody:
HRP Goat Anti- Rabbit IgG (H+L)at
1:10000 dilution.
Lysates/proteins: 25ug per lane.
Blocking buffer: 3% nonfat dry milk in
TBST.
Detection: ECL West Pico Plus
Exposure time: 10s.
Western blot analysis of extracts from
normal (control) and YAP1 knockout (KO)
HeLa cells, using YAP1 antibody at 1:1000
dilution. Secondary antibody: HRP Goat
Anti- Rabbit IgG (H+L) at 1:10000 dilution.
Lysates/proteins: 25ug per lane. Blocking
buffer: 3% nonfat dry milk in TBST.
Detection: ECL Enhanced Kit.
Exposure time: 90s.
Immunohistochemistry of paraffinembedded
human placenta using YAP1
antibody at dilution of 1:100 (40x
lens).
Immunofluorescence analysis of C6 cells
using YAP1 antibody at dilution of 1:100.
Blue: DAPI for nuclear staining.
Immunoprecipitation analysis of 200ug
extracts of HeLa cells, using 3 ug YAP1
antibody. Western blot was performed
from the immunoprecipitate using YAP1
antibody (at a dilition of 1:1000.
References
(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.