Anti-YAP1/Yap Rabbit Monoclonal Antibody

Western blot analysis of YAP1 using anti-YAP1 antibody (M00116).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human HepG2 whole cell lysates,
Lane 3: human 293T whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-YAP1 antigen affinity purified monoclonal antibody (Catalog # M00116) at 1:5000 overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:500 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for YAP1 at approximately 70 kDa. The expected band size for YAP1 is at 54 kDa.

Immunohistochemical analysis of paraffin-embedded human uterus, using YAP1 Antibody.

YAP knockdowned significantly decreased YAP expression at mRNA ( a ) and protein ( b ) levels in Hep-2 cells. c Immunofluorescence (bottom panel) was used to detect the expression of YAP (red). DAPI (blue) was used to stain the nuclei. The fluorescence intensity of YAP was stronger in YAP-NC groups, and was weaker in cells transfected with shRNA. One representative experiment out of the three performed is shown (80X). Experiments were repeated three times, and data are shown as the mean ± SD, ** P < 0.01 ( a ), ** P < 0.01 ( b )
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YAP knockdowned suppresses cell proliferation, invasion and migration. Silenced YAP inhibited cell proliferation in Hep-2 cells by CCK-8 (top panel). a The OD value at 450 nm for 5 days. Hep-2 with no treatment (Control), Non-nonspecific shRNA treatment (YAP-NC), YAP knockdown (YAP-shRNA) groups, and cDNA overexpression after YAP knockdown (YAP-shRNA+YAP-cDNA), * P < 0.05. b Colony formation assays (second panel) were performed to evaluate the proliferative capability of control, YAP-NC, YAP-shRNA and YAP-shRNA+YAP-cDNA cell groups. A representative image is shown, and a statistical comparison of the indicated groups was performed across four independent experiments, ** P < 0.01. c Wound healing assays (third panel) were performed to explore the migration capability, and solid lines represented the wound edges. Images were captured by using light microscopy (40X). The migration index was calculated as described in the Materials and Methods (control, YAP-NC, YAP-shRNA and YAP-shRNA+YAP-cDNA). Statistical analysis is shown, ** P < 0.01
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Silencing of YAP promotes cell apoptosis and induces cell cycle arrest. a Guava easyCyte12 was used to investigate differences in cell cycle (top panel) distribution following YAP silencing or overexpression in Hep-2. Silenced YAP drove G2/M arrest in control, YAP-NC, YAP-shRNA and YAP-shRNA+YAP-cDNA cell groups. In addition, when YAP knockdown Hep-2 cells overexpressed YAP, the G2/M phase distribution was decreased, ** P < 0.01; b Apoptotic cells (medium panel) were analysed by Guava easyCyte12 via staining of annexin PE/7-AAD. The percentage of apoptotic cells is shown. ** P < 0.01
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a Targeted silencing of YAP inhibits the expression of mRNA and protein levels. a qRT-PCR was used to analyse the mRNA expression of YAP, GSK-3β, DDK1, E-cadherin, β-catenin and vimentin in Hep-2 cell lines after transfection or not, and GAPDH was used as the internal control. Experiments were repeated ten times, and data are shown as the mean ± SD, ** P < 0.01. b Western blot assay was employed to investigate the expression of YAP, β-catenin, E-cadherin, vimentin, DDK1 and GSK-3β. WB results indicated that the expression of vimentin and β-catenin were downregulated with YAP knockdown and were upregulated when cells were administered YAP-cDNA. Representative images are shown. Statistical analysis of the relative optical density of each band is shown. GAPDH was used as an internal control, ** P < 0.01. c Model of the role of YAP in laryngeal cancer
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a Targeted silencing of YAP suppresses laryngeal cancer tumourigenicity and metastasis in vivo. The effects of silenced YAP on tumour suppression in vivo. Images of tumours (left panel) formed in nude mice injected subcutaneously with Hep-2 cells transfected with control, negative vector, and YAP-shRNA. Fluorescence images (right panel) of tumours captured on the IVIS system 21 days after subcutaneous injection. Cells of each group were stably transfected with luciferase. Tumours with YAP knockdown were smaller compared to the other groups. Tumour growth curves are plotted. ** P < 0.01. b A pulmonary metastasis model (left panel) was established after 3 weeks of the indicated treatment. Images from the pulmonary metastasis model and the corresponding statistical analysis are shown. ** P < 0.01
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Immunohistochemistry staining for YAP, KI-67 and Bcl-2 protein in xenograft tumour model and pulmonary metastasis model. Cytoplasmic staining was considered to be positive for YAP, KI-67 and Bcl-2. b & f Higher YAP expression in both xenograft tumour and pulmonary models. c & d High YAP expression mainly in the nucleus of LSCC. e & f Low YAP expression in LSCC in both the cytoplasm and nucleus of tumour cells. g & h Low expression of YAP protein mainly in the nucleus of LSCC. [A, a, E, e] × 40; [B, b, C, c, D, d, F, f, G, g, H, h] × 100
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Immunofluorescent analysis of MCF7 cells, using YAP1 Antibody .

Immunofluorescent analysis using the Antibody at 1:50 dilution.

Immunofluorescent analysis using the Antibody at 1:150 dilution.