CST, 14074T, YAP (D8H1X) Rabbit Monoclonal Antibody

Monoclonal Antibody for studying YAP (YAP1-2gamma). Validated for WB,WB,IP,IHC,IHC,IF,IF,F,ChIP,ChIP,C&R. Available in 3 sizes. Highly specific and rigorously validated in-house, YAP (D8H1X) Rabbit Monoclonal Antibody (CST #14074) is ready to ship. Product Usage Information For optimal ChIP and ChIP-seq results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 10 6 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 . 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 . Protocol Available protocols: Western Blotting, Immunoprecipitation, IHC Leica Bond, Immunohistochemistry (Paraffin), Immunofluorescence (Frozen), Immunofluorescence (Immunocytochemistry), Flow Cytometry (Fixed/Permeabilized), Chromatin IP, Chromatin IP-seq, CUT&RUN Specificity / Sensitivity YAP (D8H1X) Rabbit Monoclonal Antibody 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 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). Alternate Names 65 kDa Yes-associated protein; COB1; Protein yorkie homolog; Transcriptional coactivator YAP1; YAP; YAP1; YAP2; YAP65; Yes associated protein 1; Yes-associated protein 1; Yes-associated protein 1, 65kDa; yes-associated protein 2; Yes-associated protein YAP65 homolog; Yes1 associated transcriptional regulator; YKI; Yorkie homolog Specification REACTIVITY: H M R Hm Mk SENSITIVITY: Endogenous MW (kDa): 65-78 Source/Isotype: Rabbit IgG