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, rat kidney tissue. |
| Subcellular location: | Cytoplasm, Nucleus. |
| Recommended Dilutions:
WB IHC-P IF-Tissue |
1:5,000 1:2,000-1:5,000 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 with Rabbit anti-Phospho-YAP1 (S127) antibody (ET1611-69) at 1/5,000 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 1/5,000 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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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 with Rabbit anti-Phospho-YAP1 (S127) antibody (ET1611-69) at 1/5,000 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 1/5,000 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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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 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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