R Recombinant
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YAP (D8H1X) XP® Rabbit mAb #14074
Filter:
- WB
- IP
- IHC
- IF
- F
- ChIP
- C&R
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.
Supporting Data
| REACTIVITY | H M R Hm Mk |
| SENSITIVITY | Endogenous |
| MW (kDa) | 65-78 |
| Source/Isotype | Rabbit IgG |
Application Key:
- WB-Western Blotting
- IP-Immunoprecipitation
- IHC-Immunohistochemistry
- IF-Immunofluorescence
- F-Flow Cytometry
- ChIP-Chromatin Immunoprecipitation
- C&R-CUT & RUN
Species Cross-Reactivity Key:
- H-Human
- M-Mouse
- R-Rat
- Hm-Hamster
- Mk-Monkey
- Related Products
- Conjugates
Product Information
Product Usage Information
For optimal ChIP and ChIP-seq results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.
The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.
The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.
| Application | Dilution |
|---|---|
| Western Blotting | 1:1000 |
| Simple Western™ | 1:10 - 1:50 |
| Immunoprecipitation | 1:50 |
| IHC Leica Bond | 1:100 - 1:400 |
| Immunohistochemistry (Paraffin) | 1:200 - 1:800 |
| Immunofluorescence (Frozen) | 1:50 - 1:200 |
| Immunofluorescence (Immunocytochemistry) | 1:50 - 1:200 |
| Flow Cytometry (Fixed/Permeabilized) | 1:50 - 1:200 |
| Chromatin IP | 1:50 |
| Chromatin IP-seq | 1:50 |
| CUT&RUN | 1:50 |
Storage
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.
For a carrier free (BSA and azide free) version of this product see product #53921.
For a carrier free (BSA and azide free) version of this product see product #53921.
Protocol
Specificity / Sensitivity
YAP (D8H1X) XP® Rabbit mAb recognizes endogenous levels of total YAP protein. Non-specific labeling of membrane within fixed frozen mouse small intestine may be observed by immunofluorescence.
Species Reactivity:
Human, Mouse, Rat, Hamster, Monkey
The antigen sequence used to produce this antibody shares 100% sequence homology with the species listed here, but reactivity has not been tested or confirmed to work by CST. Use of this product with these species is not covered under our Product Performance Guarantee.
Species predicted to react based on 100% sequence homology:
Bovine, Horse, Guinea Pig
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.
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).
- 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.
Pathways
Explore pathways related to this product.
限制使用
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