Revision 2

#50413

Store at -20C

CST Logo
Orders:

877-616-CELL (2355)

orders@cellsignal.com

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:
W, ELISA

Reactivity:
H M R

Sensitivity:
Endogenous

MW (kDa):
65-78

Source/Isotype:
Rabbit IgG

UniProt ID:
#P46937

Entrez-Gene Id:
10413

Product Usage Information

This product is the carrier free version of product #13619. All data were generated using the same antibody clone in the standard formulation which contains BSA and glycerol.

This formulation is ideal for use with technologies requiring specialized or custom antibody labeling, including fluorophores, metals, lanthanides, and oligonucleotides. It is not recommended for ChIP, ChIP-seq, CUT&RUN or CUT&Tag assays. If you require a carrier free formulation for chromatin profiling, please contact us. Optimal dilutions/concentrations should be determined by the end user.

BSA and Azide Free antibodies are quality control tested by size exclusion chromatography (SEC) to determine antibody integrity.

Formulation

Supplied in 1X PBS (10 mM Na2HPO4, 3 mM KCl, 2 mM KH2PO4, and 140 mM NaCl (pH 7.8)). BSA and Azide Free.

For standard formulation of this product see product #13619

Storage

Store at -20°C. This product will freeze at -20°C so it is recommended to aliquot into single-use vials to avoid multiple freeze/thaw cycles. A slight precipitate may be present and can be dissolved by gently vortexing. This will not interfere with antibody performance.

Specificity/Sensitivity

Phospho-YAP (Ser397) Rabbit mAb recognizes endogenous levels of YAP protein only when phosphorylated at Ser397. This residue corresponds to Ser381 of YAP isoform 2, as reported by Zhao, B. et al. (2010) Genes Dev 24, 72-85 (9).

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser397 of human YAP protein isoform 1.

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).
Phosphorylation of YAP at Ser397 (annotated as Ser381 by Zhao et al. 2010) primes YAP for subsequent phosphorylation by CK1δ/ε in an adjacent phosphodegron (9). Upon phosphorylation, the phosphodegron recruits the β-TrCP (SCF) ubiquitin ligase complex, which ubiquitinates YAP to trigger its proteolytic degradation in the proteasome.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Applications Key

W: Western Blotting ELISA: ELISA

Cross-Reactivity Key

H: Human M: Mouse R: Rat

Trademarks and Patents

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

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Orders: 877-616-CELL (2355) orders@cellsignal.com Support: 877-678-TECH (8324) info@cellsignal.com Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 2

Cell Signaling Technology Logo
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). Data were generated using the standard formulation of this product.
Western Blotting Image 1: Phospho-YAP (Ser397) (D1E7Y) Rabbit mAb (BSA and Azide Free)
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). Data were generated using the standard formulation of this product.
Western Blotting Image 2: Phospho-YAP (Ser397) (D1E7Y) Rabbit mAb (BSA and Azide Free)
Western blot analysis of extracts from various cell lines using Phospho-YAP (Ser397) (D1E7Y) Rabbit mAb. Data were generated using the standard formulation of this product.
Western Blotting Image 3: Phospho-YAP (Ser397) (D1E7Y) Rabbit mAb (BSA and Azide Free)
Orders: 877-616-CELL (2355) orders@cellsignal.com Support: 877-678-TECH (8324) info@cellsignal.com Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 2

Cell Signaling Technology Logo
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]. Data were generated using the standard formulation of this product.
Western Blotting Image 4: Phospho-YAP (Ser397) (D1E7Y) Rabbit mAb (BSA and Azide Free)
Data using Phospho-YAP (Ser397) Matched Antibody Pair #59695 are shown. The two antibodies function as a capture-target-detection sandwich (#50413 and #27166, respectively) to detect endogenous protein expression in positive and negative controls.
ELISA Image 1: Phospho-YAP (Ser397) (D1E7Y) Rabbit mAb (BSA and Azide Free)
Orders: 877-616-CELL (2355) orders@cellsignal.com Support: 877-678-TECH (8324) info@cellsignal.com Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.