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Phospho-YAP1 (S127) Recombinant Rabbit Monoclonal Antibody [SN0718]

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Specification

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Catalog# ET1611-69

Phospho-YAP1 (S127) Recombinant Rabbit Monoclonal Antibody [SN0718]

Application

  • WB

  • IHC-P

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

Phospho-YAP1 (S127) Recombinant Rabbit Monoclonal Antibody [SN0718]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic phospho-peptide corresponding to residues surrounding Ser127 of human YAP1.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P

Target Molecular Weight

Predicted band size: 54 kDa

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, human kidney tissue, mouse kidney tissue, rat kidney tissue.

Conjugation

unconjugated

Clone Number

SN0718

RRID

AB_3070048

Product Features

Form

Liquid

Concentration

1 mg/mL.(The concentration of this product may be batch-dependent)

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:5,000

  • IHC-P

  • 1:2,000-1:5,000

Target

Function

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.

Background References

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).

Sequence Similarity

Belongs to the YAP1 family.

Tissue Specificity

Increased expression seen in some liver and prostate cancers. Isoforms lacking the transactivation domain found in striatal neurons of patients with Huntington disease (at protein level).

Post-translational Modification

Phosphorylated by LATS1 and LATS2; leading to cytoplasmic translocation and inactivation. Phosphorylated by ABL1; leading to YAP1 stabilization, enhanced interaction with TP73 and recruitment onto proapoptotic genes; in response to DNA damage. Phosphorylation at Ser-400 and Ser-403 by CK1 is triggered by previous phosphorylation at Ser-397 by LATS proteins and leads to YAP1 ubiquitination by SCF(beta-TRCP) E3 ubiquitin ligase and subsequent degradation. Phosphorylated at Thr-119, Ser-138, Thr-154, Ser-367 and Thr-412 by MAPK8/JNK1 and MAPK9/JNK2, which is required for the regulation of apoptosis by YAP1.; Ubiquitinated by SCF(beta-TRCP) E3 ubiquitin ligase.

Subcellular Location

Cytoplasm, Nucleus.

UNIPROT

SwissProt: P46937 Human

SwissProt: P46938 Mouse

SwissProt: Q2EJA0 Rat

Synonyms

65 kDa Yes associated protein antibody

65 kDa Yes-associated protein antibody

COB1 antibody

YAp 1 antibody

YAP 65 antibody

YAP antibody

YAP1 antibody

YAP1_HUMAN antibody

YAP2 antibody

YAP65 antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Phospho-YAP1 (S127) on different lysates with Rabbit anti-Phospho-YAP1 (S127) antibody (<a href="/products/ET1611-69" style="font-weight: bold;text-decoration: underline;">ET1611-69</a>) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: HeLa treated with 100nM Calyculin A for 30 minutes cell lysate<br />Lane 3: C6 cell lysate<br />Lane 4: C6 treated with 100nM Calyculin A for 30 minutes cell lysate<br />Lane 5: NIH/3T3 cell lysate<br />Lane 6: NIH/3T3 starved for 24 hours then treated with 100nM Calyculin A for 30 minutes cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 54 kDa<br />Observed band size: 70 kDa<br /><br />Exposure time: 50 seconds; ECL: K1802;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1611-69" style="font-weight: bold;text-decoration: underline;">ET1611-69</a>) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution were used in 5% BSA at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    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 cell lysate
    Lane 6: NIH/3T3 starved for 24 hours then treated with 100nM Calyculin A for 30 minutes cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 54 kDa
    Observed band size: 70 kDa

    Exposure time: 50 seconds; ECL: K1802;

    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% BSA at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Phospho-YAP1 (S127) on different lysates with Rabbit anti-Phospho-YAP1 (S127) antibody (<a href="/products/ET1611-69" style="font-weight: bold;text-decoration: underline;">ET1611-69</a>) at 1/5,000 dilution.<br /><br />Lane 1: HeLa whole cell lysate<br />Lane 2: HeLa treated with 100nM Calyculin A for 30 minutes whole cell lysate<br />Lane 3: HeLa treated with 100nM Calyculin A for 30 minutes then treated with λpp for 1 hour whole cell lysate<br />Lane 4: NIH/3T3 whole cell lysate<br />Lane 5: NIH/3T3 treated with 100nM Calyculin A for 30 minutes whole cell lysate<br />Lane 6: NIH/3T3 treated with 100nM Calyculin A for 30 minutes then treated with λpp for 1 hour whole cell lysate<br />Lane 7: C6 whole cell lysate<br />Lane 8: C6 treated with 100nM Calyculin A for 30 minutes whole cell lysate<br />Lane 9: C6 treated with 100nM Calyculin A for 30 minutes then treated with λpp for 1 hour whole cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 54 kDa<br />Observed band size: 70 kDa<br /><br />Exposure time: 1 minute 6 seconds;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1611-69" style="font-weight: bold;text-decoration: underline;">ET1611-69</a>) at 1/5,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:100,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    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.

  • Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Phospho-YAP1 (S127) antibody (<a href="/products/ET1611-69" style="font-weight: bold;text-decoration: underline;">ET1611-69</a>) at 1/5,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1611-69" style="font-weight: bold;text-decoration: underline;">ET1611-69</a>) 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.

    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.

  • Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-Phospho-YAP1 (S127) antibody (<a href="/products/ET1611-69" style="font-weight: bold;text-decoration: underline;">ET1611-69</a>) at 1/5,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1611-69" style="font-weight: bold;text-decoration: underline;">ET1611-69</a>) 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.

    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.

  • Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-Phospho-YAP1 (S127) antibody (<a href="/products/ET1611-69" style="font-weight: bold;text-decoration: underline;">ET1611-69</a>) at 1/2,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1611-69" style="font-weight: bold;text-decoration: underline;">ET1611-69</a>) 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.

    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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Citation