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PathScan® Total YAP Sandwich ELISA Kit #32939

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Figure 1: YAP protein is expressed in a variety of human cells but absent in the Jurkat cell line, as detected by using the PathScan® Total YAP Sandwich ELISA Kit. The absorbance readings at 450 nm are shown in the top figure, while the corresponding Western blot using YAP (1A12) Mouse mAb #12395 is shown in the bottom figure.

To Purchase # 32939**

Important Ordering Details

Custom Ordering Details:

If kit quantities from the same lot are needed in unlisted sizes, contact us for processing time and pricing.

Looking for this ELISA kit in a 384-well format? Inquire for availability, processing time, and pricing.

Supporting Data

REACTIVITY H M
Application Key:
  • ELISA-ELISA 
Species Cross-Reactivity Key:
  • H-Human 
  • M-Mouse 
  • Product Includes
  • Related Products
Product IncludesVolumeSolution Color
YAP Rabbit mAb Coated Microwells #7996396 tests
YAP Mouse Detection mAb #430061 eaGreen (Lyophilized)
Anti-mouse IgG, HRP-linked Antibody (ELISA Formulated) #133041 eaRed (Lyophilized)
Detection Antibody Diluent #1333911 mlGreen
HRP Diluent #1351511 mlRed
TMB Substrate #700411 ml
STOP Solution #700211 ml
Sealing Tape #545032 ea
ELISA Wash Buffer (20X) #980125 ml
ELISA Sample Diluent #1108325 mlBlue
Cell Lysis Buffer (10X) #980315 ml

Kit contents scale proportionally with size, except sealing tape.
Example: The V1 kit contains 5X the listed quantities above, but will exclude the sealing tape.

The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.

Product Information

Product Description

The PathScan® Total YAP Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of YAP protein. A YAP rabbit mAb has been coated onto the microwells. After incubation with cell lysates, the YAP protein is captured by the coated antibody. Following extensive washing, YAP mouse detection mAb is added to detect captured YAP protein. Anti-mouse, HRP-linked antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of absorbance for the developed color is proportional to the quantity of YAP protein.

*Antibodies in this kit are custom formulations specific to kit.

Protocol

Specificity / Sensitivity

PathScan® Total YAP Sandwich ELISA Kit detects endogenous levels of YAP protein in human cells, as shown in Figure 1. The kit sensitivity is shown in Figure 2. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Species Reactivity:

Human, Mouse

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).

Pathways

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