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YAP1 Recombinant Rabbit Monoclonal Antibody [SU33-06]

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Specification

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Catalog# ET1608-30

YAP1 Recombinant Rabbit Monoclonal Antibody [SU33-06]

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • IHC-P

  • IP

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

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

Overview

Product Name

YAP1 Recombinant Rabbit Monoclonal Antibody [SU33-06]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human YAP1 aa 421-470 / 504.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IF-Tissue, IHC-P, IP, FC

Molecular Weight

Predicted band size: 54 kDa

Positive Control

PC-12 cell lysate, rat liver tissue lysate, HeLa, human colon carcinoma tissue, human breast carcinoma tissue, human kidney tissue, HCT 116 cell lysate, Mouse liver tissue lysate, Mouse kidney tissue lysate, Mouse testis tissue lysate, Mouse skin tissue lysate.

Conjugation

unconjugated

Clone Number

SU33-06

RRID

AB_3069799

Product Features

Form

Liquid

Concentration

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

  • 1:1,000-1:2,000

  • IF-Cell

  • 1:100

  • IF-Tissue

  • 1:200

  • IHC-P

  • 1:200

  • FC

  • 1:1,000

  • IP

  • Use at an assay dependent concentration.

Target

Function

YAP1 (yes-associated protein 1), also known as YAP or YAP65, is a protein that acts as a transcription coregulator that promotes transcription of genes involved in cellular proliferation and suppressing apoptotic genes. YAP1 is a component in the hippo signaling pathway which regulates organ size, regeneration, and tumorigenesis. YAP1 was first identified by virtue of its ability to associate with the SH3 domain of Yes and Src protein tyrosine kinases. YAP1 is a potent oncogene, which is amplified in various human cancers.

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

  • Western blot analysis of YAP1 on different lysates with Rabbit anti-YAP1 antibody (<a href="/products/ET1608-30" style="font-weight: bold;text-decoration: underline;">ET1608-30</a>) at 1/1,000 dilution.<br /><br />Lane 1: PC-12 cell lysate (10 µg/Lane)<br />Lane 2: Rat liver tissue lysate (20 µg/Lane)<br /><br />Predicted band size: 54 kDa<br />Observed band size: 75 kDa<br /><br />Exposure time: 1 minute;<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/ET1608-30" style="font-weight: bold;text-decoration: underline;">ET1608-30</a>) at 1/1,000 dilution was used in 5% NFDM/TBST 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.

    Western blot analysis of YAP1 on different lysates with Rabbit anti-YAP1 antibody (ET1608-30) at 1/1,000 dilution.

    Lane 1: PC-12 cell lysate (10 µg/Lane)
    Lane 2: Rat liver tissue lysate (20 µg/Lane)

    Predicted band size: 54 kDa
    Observed band size: 75 kDa

    Exposure time: 1 minute;

    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 (ET1608-30) at 1/1,000 dilution was 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.

  • Western blot analysis of YAP1 on different lysates with Rabbit anti-YAP1 antibody (<a href="/products/ET1608-30" style="font-weight: bold;text-decoration: underline;">ET1608-30</a>) at 1/5,000 dilution.<br /><br />Lane 1: HepG2 cell lysate<br />Lane 2: Hela cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br />Exposure time: 25 seconds; ECL: K1801<br /><br />Blocking: 5% NFDM/TBST, 1 hour at room temperature<br />Primary antibody: <a href="/products/ET1608-30" style="font-weight: bold;text-decoration: underline;">ET1608-30</a>, 1/5,000 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), overnight at 4 ℃<br />Secondary antibody: Goat anti-Rabbit IgG-HRP (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature<br /><br />Predicted band size: 54kDa<br />Observed band size: 70kDa

    Western blot analysis of YAP1 on different lysates with Rabbit anti-YAP1 antibody (ET1608-30) at 1/5,000 dilution.

    Lane 1: HepG2 cell lysate
    Lane 2: Hela cell lysate

    Lysates/proteins at 20 µg/Lane.
    Exposure time: 25 seconds; ECL: K1801

    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: ET1608-30, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
    Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

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

  • Immunocytochemistry analysis of HeLa cells labeling YAP1 with Rabbit anti-YAP1 antibody (<a href="/products/ET1608-30" style="font-weight: bold;text-decoration: underline;">ET1608-30</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-YAP1 antibody (<a href="/products/ET1608-30" style="font-weight: bold;text-decoration: underline;">ET1608-30</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of HeLa cells labeling YAP1 with Rabbit anti-YAP1 antibody (ET1608-30) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-YAP1 antibody (ET1608-30) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Flow cytometric analysis of HeLa cells labeling YAP1.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/ET1608-30" style="font-weight: bold;text-decoration: underline;">ET1608-30</a>, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of HeLa cells labeling YAP1.

    Cells were fixed and permeabilized. Then stained with the primary antibody (ET1608-30, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

  • Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-YAP1 antibody (<a href="/products/ET1608-30" style="font-weight: bold;text-decoration: underline;">ET1608-30</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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/ET1608-30" style="font-weight: bold;text-decoration: underline;">ET1608-30</a>) at 1/200 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 colon carcinoma tissue with Rabbit anti-YAP1 antibody (ET1608-30) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (ET1608-30) at 1/200 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 breast carcinoma tissue with Rabbit anti-YAP1 antibody (<a href="/products/ET1608-30" style="font-weight: bold;text-decoration: underline;">ET1608-30</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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/ET1608-30" style="font-weight: bold;text-decoration: underline;">ET1608-30</a>) at 1/200 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 breast carcinoma tissue with Rabbit anti-YAP1 antibody (ET1608-30) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (ET1608-30) at 1/200 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-YAP1 antibody (<a href="/products/ET1608-30" style="font-weight: bold;text-decoration: underline;">ET1608-30</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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/ET1608-30" style="font-weight: bold;text-decoration: underline;">ET1608-30</a>) at 1/200 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-YAP1 antibody (ET1608-30) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (ET1608-30) at 1/200 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.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of YAP1 on different lysates with Rabbit anti-YAP1 antibody (<a href="/products/ET1608-30" style="font-weight: bold;text-decoration: underline;">ET1608-30</a>) at 1/1,000 dilution.<br /><br />Lane 1: HCT 116-si NT cell lysate<br />Lane 2: HCT 116-si YAP1 cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 54 kDa<br />Observed band size: 75 kDa<br /><br />Exposure time: 20 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/ET1608-30" style="font-weight: bold;text-decoration: underline;">ET1608-30</a>) at 1/1,000 dilution was 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.

    ☑ Knockdown (KD)

    Western blot analysis of YAP1 on different lysates with Rabbit anti-YAP1 antibody (ET1608-30) at 1/1,000 dilution.

    Lane 1: HCT 116-si NT cell lysate
    Lane 2: HCT 116-si YAP1 cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 54 kDa
    Observed band size: 75 kDa

    Exposure time: 20 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 (ET1608-30) at 1/1,000 dilution was 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.

  • Western blot analysis of YAP1 on different lysates with Rabbit anti-YAP1 antibody (<a href="/products/ET1608-30" style="font-weight: bold;text-decoration: underline;">ET1608-30</a>) at 1/1,000 dilution.<br /><br />Lane 1: Mouse liver tissue lysate<br />Lane 2: Mouse kidney tissue lysate<br />Lane 3: Mouse testis tissue lysate<br />Lane 4: Mouse skin tissue lysate<br /><br />Lysates/proteins at 40 µg/Lane.<br /><br />Predicted band size: 54 kDa<br />Observed band size: 70 kDa<br /><br />Exposure time: 2 minutes; 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/ET1608-30" style="font-weight: bold;text-decoration: underline;">ET1608-30</a>) at 1/1,000 dilution was used in 5% NFDM/TBST 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.

    Western blot analysis of YAP1 on different lysates with Rabbit anti-YAP1 antibody (ET1608-30) at 1/1,000 dilution.

    Lane 1: Mouse liver tissue lysate
    Lane 2: Mouse kidney tissue lysate
    Lane 3: Mouse testis tissue lysate
    Lane 4: Mouse skin tissue lysate

    Lysates/proteins at 40 µg/Lane.

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

    Exposure time: 2 minutes; 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 (ET1608-30) at 1/1,000 dilution was 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.

  • Application: IF-Tissue<br /><br />Species: Human<br /><br />Site: colon carcinoma<br /><br />Sample: Paraffin-embedded section<br /><br />Antibody concentration: 1/200

    Application: IF-Tissue

    Species: Human

    Site: colon carcinoma

    Sample: Paraffin-embedded section

    Antibody concentration: 1/200

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation