产品描述The Yes-associated protein, otherwise known as YAP, is a 14-3-3-binding molecule that was originally recognized by virtue of its ability to bind to the SH3 domain of Yes. The binding of YAP to 14-3-3 requires the phosphorylation of a homologous serine residue (Ser 112) in the YAP 14-3-3-binding motif. The highly conserved and ubiquitously expressed 14-3-3 proteins regulate differentiation, cell cycle progression and apoptosis by binding intracellular phosphoproteins involved in signal transduction. YAP may link events at the plasma membrane and cytoskeleton to inhibition of transcription in the nucleus in a manner regulated by 14-3-3 proteins. YAP shares homology with the WW domain of TAZ, transcriptional co-activator with PDZ-binding motif, which functions as a transcriptional co-activator by binding to the PPXY motif present in transcription factors. YAP is expressed at high levels in the lung, placenta, prostate, ovary and testis.
Fig1: Western blot analysis of Phospho-YAP1 (S127) on SiHa cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1611-69, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
Fig2: Western blot analysis of Phospho-YAP1 (S127) on SiHa cell lysates.
Lane 1: SiHa cells, whole cell lysate, 10ug/lane
Lane 2 : SiHa cells treated with 2.8ug/ul lambda-PP for 30 minutes, whole cell lysates, 10ug/lane
All lanes :
Anti-Phospho-YAP1 (S127) antibody (ET1611-69) at 1/500 dilution. Anti-YAP1 (S127) antibody (ET1608-30) at 1/500 dilution. Anti-GAPDH antibody (ET1601-4) at 1/10,000 dilution. Goat Anti-Rabbit IgG H&L (HRP) (HA1001) at 1/200,000 dilution.
Predicted band size: 54 kDa
Observed band size: 70 kDa
Blocking and diluting buffer: 5% BSA.
Exposure time: 2 minutes 34 seconds
Fig3: Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-Phospho-YAP1 (S127) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1611-69, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Fig4: Immunohistochemical analysis of paraffin-embedded mouse kidney tissue using anti-Phospho-YAP1 (S127) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1611-69, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
背景文献
1. Ye L et al. Decreased Yes-Associated Protein-1 (YAP1) Expression in Pediatric Hearts with Ventricular Septal Defects. PLoS One 10:e0139712 (2015).
2. Whiteman EL et al. Crumbs3 is essential for proper epithelial development and viability. Mol Cell Biol 34:43-56 (2014).
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