Overview

  • Product name
    Anti-active YAP1 antibody [EPR19812]
    See all active YAP1 primary antibodies
  • Description
    Rabbit monoclonal [EPR19812] to active YAP1
  • Host species
    Rabbit
  • Specificity
    ab205270 is specific to the active (non-phosphorylated) form of YAP1.
  • Tested applications
    Suitable for: ICC/IF, IHC-P, WBmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    This product was produced with the following immunogens:
    Synthetic peptide within Human active YAP1 aa 100-200. The exact sequence is proprietary.
    Database link: P46937

    Synthetic peptide within Human active YAP1 aa 100-200. The exact sequence is proprietary.
    Database link: P46937

  • Positive control
    • WB: 293A cell lysate serum starved overnight, then 10% FBS was added to medium for 1 hour; 293A cell lysate serum starved overnight and then treated with Lambda phosphatase lysate. Human kidney and skin lysates and mouse testis and skin lysates. HaCaT whole cell lysate, 293A cell lysate. IHC-P: Human breast and breast cancer tissues; Mouse skin tissue. ICC/IF: 293A cell line serum starved overnight, then 10% FBS was added to medium for 1 hour; 293A cells.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab205270 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF 1/500.
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/1000. Detects a band of approximately 75 kDa (predicted molecular weight: 54 kDa).

Target

Images

  • All lanes : Anti-active YAP1 antibody [EPR19812] (ab205270) at 1 µg/ml

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : active YAP1 knockout HAP1 whole cell lysate
    Lane 3 : HeLa whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 54 kDa



    Lanes 1 - 3: Merged signal (red and green). Green - ab205270 observed at 54 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab205270 was shown to recognize active AP1 in wild-type HAP1 cells as signal was lost at the expected MW in active YAP1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and active YAP1 knockout samples were subjected to SDS-PAGE. Ab205270 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemical analysis of paraffin-embedded human breast tissue labeling active YAP1 with ab205270 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Mainly nuclear staining on human breast is observed [PMID: 18617895].

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Ab205270 staining active YAP1 in HUVEC (human umbilical vein endothelial cell) cells by Immunocytochemistry/Immunofluorescence (ICC/IF). The cells were fixed 4% Paraformaldehyde and permeabilized with 0.1% TritonX-100. Samples were incubated with primary antibody at 1:500 dilution. An Alexa Fluor® 488 Goat anti-Rabbit was used as a secondary antibody at 1:1000 dilution. DAPI was used as a nuclear counter stain. Confocal image showing nuclear and cytoplasmic staining in HUVEC cells.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton/PBS permeabilized (RT, 5 mins) 293A (Human epithelial cell line from embryonic kidney transformed with sheared human adenovirus type 5 DNA) cells labeling active YAP1 with ab205270 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 594) secondary antibody at 1/1000 dilution.

    The images showed weak staining on 293A cells under serum starvation overnight. After 10% FBS was added to the medium for 1h, the nuclear staining was increased.

    The data was kindly provided by our collaborator Dr. Bin Zhao (Zhejiang University).

    The nuclear counterstain is DAPI (blue). Counterstained with Phalloidin-technology® Alexa Fluor 488 at 1/1000 dilution.

  • All lanes : Anti-active YAP1 antibody [EPR19812] (ab205270) at 1/1000 dilution

    Lane 1 : 293A cells under serum starvation overnight lysate
    Lane 2 : 293A cell lysate under serum starvation overnight, then 10% FBS was added to medium for 1 hour

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 54 kDa
    Observed band size: 75 kDa
    why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Serum starvation induces active YAP1 Ser127 phosphorylation. The level of active YAP1 protein is inversely proportional to p-YAP1 Ser127 level (PMID: 22884261).

  • All lanes : Anti-active YAP1 antibody [EPR19812] (ab205270) at 1/1000 dilution

    Lane 1 : Human kidney lysate
    Lane 2 : Human skin lysate
    Lane 3 : HaCaT (Human keratinocyte cell line) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Lanes 1-2 : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution
    Lane 3 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 54 kDa
    Observed band size: 75 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1/2: 8 seconds; Lane 3: 30 seconds.

  • All lanes : Anti-active YAP1 antibody [EPR19812] (ab205270) at 1/1000 dilution

    Lane 1 : Mouse testis lysate
    Lane 2 : Mouse skin lysate
    Lane 3 : Mouse liver lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 54 kDa
    Observed band size: 75 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1: 10 seconds; Lane 2: 8 seconds; Lane 3: 3 minutes.

  • All lanes : Anti-active YAP1 antibody [EPR19812] (ab205270) at 1/1000 dilution

    Lane 1 : 293A cell lysate serum starved overnight and then 10% FBS added to the medium for 1 hour
    Lane 2 : 293A cell lysate serum starved overnight
    Lane 3 : 293A cell lysate serum starved overnight and then treated with Lambda phosphatase

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : donkey anti-rabbit at 1/1000 dilution

    Predicted band size: 54 kDa
    Observed band size: 75 kDa why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    Blocking/Dilution buffer: 5% BSA/TBST.

    The two panels for pYAPS127 were just shorter and longer exposure.

    The data was kindly provided by our collaborator Dr. Bin Zhao (Zhejiang University).

  • All lanes : Anti-active YAP1 antibody [EPR19812] (ab205270) at 1/1000 dilution

    Lane 1 : 293A cell lysate
    Lane 2 : YAP/TAZ knockout 293A cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : donkey anti-rabbit at 1/1000 dilution

    Predicted band size: 54 kDa
    Observed band size: 75 kDa why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    Blocking/Dilution buffer: 5% BSA/TBST.

    The data was kindly provided by our collaborator Dr. Bin Zhao (Zhejiang University).

  • Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling active YAP1 with ab205270 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Nuclear and cytoplasmic staining on human breast cancer is observed [PMID: 24559095].

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunohistochemical analysis of paraffin-embedded mouse skin tissue labeling active YAP1 with ab205270 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Mainly nuclear staining on mouse skin is observed [PMID: 21610251].

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton/PBS permeabilized (RT, 5 mins) 293A (Human epithelial cell line from embryonic kidney transformed with sheared human adenovirus type 5 DNA) cells labeling active YAP1 with ab205270 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 594) secondary antibody at 1/1000 dilution.

    The images showed nuclear staining on 293A cells, and background staining on YAP/TAZ knockout 293A cells.

    The data was kindly provided by our collaborator Dr. Bin Zhao (Zhejiang University).

    The nuclear counterstain is DAPI (blue). Counterstained with Phalloidin-technology® Alexa Fluor 488 at 1/1000 dilution.

  • Immunohistochemical analysis of paraffin-embedded human liver tissue labeling active YAP1 with ab205270 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Negative control: no staining on human liver [PMID:17974916].

    Counter stained with Hematoxylin.

References

This product has been referenced in:
  • Turunen SP  et al. FGFR4 phosphorylates MST1 to confer breast cancer cells resistance to MST1/2-dependent apoptosis. Cell Death Differ N/A:N/A (2019). Read more (PubMed: 30903103) »
  • Lodge EJ  et al. Homeostatic and tumourigenic activity of SOX2+ pituitary stem cells is controlled by the LATS/YAP/TAZ cascade. Elife 8:N/A (2019). Read more (PubMed: 30912742) »
See all 7 Publications for this product

Customer reviews and Q&As

1-10 of 11 Abreviews or Q&A

Application
IHC - Wholemount
Sample
Mouse Embryo (E14.5 skin)
Specification
E14.5 skin

Dr. Vinod Kumar

Verified customer

Submitted Nov 20 2018

Application
Immunoprecipitation
Sample
Mouse Tissue lysate - other (kidney cells)
Total protein in input
900 µg
Immuno-precipitation step
Protein A
Specification
kidney cells

Abcam user community

Verified customer

Submitted Oct 18 2018

Application
Western blot
Sample
Mouse Cell lysate - whole cell (mouse neural stem cell)
Gel Running Conditions
Reduced Denaturing (4-12%)
Loading amount
50 µg
Specification
mouse neural stem cell
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Dr. Robert Kupp

Verified customer

Submitted Sep 05 2018

Application
IHC - Wholemount
Sample
Mouse Tissue (intestine organoids)
Specification
intestine organoids

Abcam user community

Verified customer

Submitted Jul 11 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Kidney)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Antigen Retrieval Buffer (100X Citrate Buffer pH 6.0) (ab94674)
Permeabilization
Yes - 0.05% tween 20
Specification
Kidney
Blocking step
Sea Block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 22°C
Fixative
Formaldehyde

Mr. David Ivancic

Verified customer

Submitted Oct 30 2017

Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (lung)
Permeabilization
Yes - 80% Acetone
Specification
lung
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Sep 29 2017

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (stomach)
Permeabilization
Yes - 0.05% TX-100
Specification
stomach
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Aug 28 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Sheep Tissue sections (Artery)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: TE pH9
Permeabilization
No
Specification
Artery
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
10% Buffered Normal Formalin

Abcam user community

Verified customer

Submitted Aug 23 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Pig Tissue sections (lung)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: TE pH9
Permeabilization
No
Specification
lung
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
10% buffered normal formalin

Abcam user community

Verified customer

Submitted Aug 21 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (tumor)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: TE pH9
Permeabilization
No
Specification
tumor
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
10% normal buffered formalin

Abcam user community

Verified customer

Submitted Aug 04 2017

1-10 of 11 Abreviews or Q&A

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