Phospho-YAP1 (S127) Recombinant Rabbit Monoclonal Antibody [SN0718]
cat.: ET1611-69
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IHC-P, IF-Tissue |
| Clonality: | Monoclonal |
| Clone number: | SN0718 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 54 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic phospho-peptide corresponding to residues surrounding Ser127 of human YAP1. |
| Positive control: | HeLa treated with 100nM Calyculin A for 30 minutes whole cell lysate, NIH/3T3 treated with 100nM Calyculin A for 30 minutes whole cell lysate, C6 treated with 100nM Calyculin A for 30 minutes whole cell lysate, SiHa cell lysates, human kidney tissue, mouse kidney tissue. |
| Subcellular location: | Cytoplasm, Nucleus. |
| Recommended Dilutions:
WB IHC-P IF-Tissue |
1:5,000 1:50-1:200 1:50-1:400 |
| Uniprot #: | SwissProt: P46937 Human | P46938 Mouse | Q2EJA0 Rat |
| Alternative names: | 65 kDa Yes associated protein 65 kDa Yes-associated protein COB1 YAp 1 YAP 65 YAP YAP1 YAP1_HUMAN YAP2 YAP65 yes -associated protein delta Yes associated protein 1 65kDa Yes associated protein 1 Yes associated protein 2 yes associated protein beta YKI Yorkie homolog |
Images
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Fig1:
Western blot analysis of Phospho-YAP1 (S127) on different lysates with Rabbit anti-Phospho-YAP1 (S127) antibody (ET1611-69) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution. Lane 1: HeLa cell lysate Lane 2: HeLa treated with 100nM Calyculin A for 30 minutes cell lysate Lane 3: C6 cell lysate Lane 4: C6 treated with 100nM Calyculin A for 30 minutes cell lysate Lane 5: NIH/3T3 starved for 24 hours then treated with 100nM Calyculin A for 30 minutes cell lysate Lane 6: NIH/3T3 cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 54 kDa Observed band size: 70 kDa Exposure time: 50 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1611-69) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig2: 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. |
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Fig3:
Western blot analysis of Phospho-YAP1 (S127) on different lysates with Rabbit anti-Phospho-YAP1 (S127) antibody (ET1611-69) at 1/5,000 dilution. Lane 1: HeLa whole cell lysate Lane 2: HeLa treated with 100nM Calyculin A for 30 minutes whole cell lysate Lane 3: HeLa treated with 100nM Calyculin A for 30 minutes then treated with λpp for 1 hour whole cell lysate Lane 4: NIH/3T3 whole cell lysate Lane 5: NIH/3T3 treated with 100nM Calyculin A for 30 minutes whole cell lysate Lane 6: NIH/3T3 treated with 100nM Calyculin A for 30 minutes then treated with λpp for 1 hour whole cell lysate Lane 7: C6 whole cell lysate Lane 8: C6 treated with 100nM Calyculin A for 30 minutes whole cell lysate Lane 9: C6 treated with 100nM Calyculin A for 30 minutes then treated with λpp for 1 hour whole cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 54 kDa Observed band size: 70 kDa Exposure time: 1 minute 6 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1611-69) at 1/5,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature. |
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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. |
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Fig5:
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 |
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Fig6:
Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-Phospho-YAP1 (S127) antibody (ET1611-69) at 1/2,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1611-69) at 1/1,000 dilution for 1 hour 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. |
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