Astana Biomed Group, an authorized Abcam distributor in Central Asia

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

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR19812] to active YAP1 - BSA and Azide free
  • Suitable for: ICC/IF, WB, IHC-P
  • Knockout validated
  • Reacts with: 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. This information is proprietary to Abcam and/or its suppliers.

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • 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

    ab223126 is the carrier-free version of ab205270.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    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

Images

  • Western blot - Anti-active YAP1 antibody [EPR19812] - BSA and Azide free (ab223126)
    Western blot - Anti-active YAP1 antibody [EPR19812] - BSA and Azide free (ab223126)
    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).

  • Immunocytochemistry/ Immunofluorescence - Anti-active YAP1 antibody [EPR19812] - BSA and Azide free (ab223126)
    Immunocytochemistry/ Immunofluorescence - Anti-active YAP1 antibody [EPR19812] - BSA and Azide free (ab223126)

    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).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-active YAP1 antibody [EPR19812] - BSA and Azide free (ab223126)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-active YAP1 antibody [EPR19812] - BSA and Azide free (ab223126)

    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).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-active YAP1 antibody [EPR19812] - BSA and Azide free (ab223126)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-active YAP1 antibody [EPR19812] - BSA and Azide free (ab223126)

    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).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-active YAP1 antibody [EPR19812] - BSA and Azide free (ab223126)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-active YAP1 antibody [EPR19812] - BSA and Azide free (ab223126)

    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).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-active YAP1 antibody [EPR19812] - BSA and Azide free (ab223126)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-active YAP1 antibody [EPR19812] - BSA and Azide free (ab223126)

    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).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-active YAP1 antibody [EPR19812] - BSA and Azide free (ab223126)
    Immunocytochemistry/ Immunofluorescence - Anti-active YAP1 antibody [EPR19812] - BSA and Azide free (ab223126)

    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).

  • Immunocytochemistry/ Immunofluorescence - Anti-active YAP1 antibody [EPR19812] - BSA and Azide free (ab223126)
    Immunocytochemistry/ Immunofluorescence - Anti-active YAP1 antibody [EPR19812] - BSA and Azide free (ab223126)

    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).

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

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