Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-active YAP1 antibody [EPR19812] - BSA and Azide free (ab223126)

Knockout Tested Rabbit recombinant monoclonal active YAP1 antibody [EPR19812]. Validated in WB, IHC, ICC/IF and tested in Mouse, Human.

Overview

  • Product name

    Anti-active YAP1 antibody [EPR19812] - BSA and Azide free
    See all active YAP1 primary antibodies
  • Description

    Rabbit monoclonal [EPR19812] to active YAP1 - BSA and Azide free
  • Host species

    Rabbit
  • Specificity

    ab223126 is specific to the active (non-phosphorylated) form of YAP1.
  • Tested applications

    Suitable for: ICC/IF, WB, IHC-Pmore 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

    The formulation and the concentration of this product is compatible for metal-conjugation for mass cytometry (CyTOF®).

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

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

Properties

Applications

Our Abpromise guarantee covers the use of ab223126 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 Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 75 kDa (predicted molecular weight: 54 kDa).
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target

Images

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

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : 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 YAP1 in wild-type HAP1 cells as signal was lost at the expected MW in YAP1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab205270).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab205270).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab205270).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab205270).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab205270).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab205270).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab205270).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab205270).

References

ab223126 has not yet been referenced specifically in any publications.

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