Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (ab51067)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP1454Y] to Met (c-Met) - N-terminal
  • Suitable for: ELISA, WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human, Recombinant fragment

Overview

  • Product name

    Anti-Met (c-Met) antibody [EP1454Y] - N-terminal
    See all Met (c-Met) primary antibodies

  • Description

    Rabbit monoclonal [EP1454Y] to Met (c-Met) - N-terminal

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    ELISA
    Recombinant fragment
    ICC/IF
    Human
    IHC-P
    Human
    WB
    Mouse
    Rat
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide within Human Met (c-Met) aa 1-100 (N terminal). The exact sequence is proprietary.
    Database link: P08581
    (Peptide available as ab167073)

  • Positive control

    • WB: Wild-type HAP1 cell lysate. HepG2, HEK-293 and HeLa cell lysate. Mouse and rat thymus tissue lysate. Mouse lung tissue lysate. Hela and A459 whole cell lysate. IHC-P: Human bladder carcinoma and clear cell kidney carcinoma tissue. ICC/IF: Jurkat cells.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

Images

  • Western blot - Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (ab51067)
    Western blot - Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (ab51067)
    All lanes : Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (ab51067) at 1/1000 dilution

    Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
    Lane 2 : A549 (human lung carcinoma epithelial cell) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size: 156 kDa
    Observed band size: 240 kDa
    why is the actual band size different from the predicted?



    Low expression:A549 (PMID: 32792859).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (ab51067)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (ab51067)

    Immunohistochemical staining of paraffin embedded human bladder carcinoma with purified ab51067 at a working dilution of 1/100. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counterstained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0.

    PBS was used instead of the primary antibody as the negative control (inset).

  • Western blot - Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (ab51067)
    Western blot - Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (ab51067)
    All lanes : Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (ab51067) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : MET knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 156 kDa
    Observed band size: 160 kDa why is the actual band size different from the predicted?



    Lanes 1 - 2: Merged signal (red and green). Green - ab51067 observed at 156 kDa. Red - loading control, ab8245 observed at 37 kDa.  

     ab51067 was shown to react with Met (c-Met) in wild-type HeLa. Loss of signal was observed when knockout cell line ab265961 (knockout cell lysate ab256991) was used. Wild-type and Met (c-Met) knockout samples were subjected to SDS-PAGE. ab51067 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

     

     

  • Western blot - Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (ab51067)
    Western blot - Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (ab51067)

    Lane 1: Wild-type HAP1 cell lysate (40 µg)
    Lane 2: Met (c-Met) knockout HAP1 cell lysate (40 µg)
    Lane 3: HepG2 cell lysate (40µg) (40 µg)
    Lane 4: HEK-293 cel lysate (40µg) (40 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab51067 observed at 240 kDa. Red - loading control, ab18058, observed at 124 kDa.

    This western blot image is a comparison between ab51067 and a competitor's top cited rabbit polyclonal antibody.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (ab51067)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (ab51067) This image is courtesy of an Abreview submitted by David Ivancic.

    ab51067 staining Met (c-Met) in human breast tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections).

    Tissue was fixed with formaldehyde, permeabilized with 0.05% Tween-20 and blocked for 30 minutes at 22°C; antigen retrieval was by heat mediation in antigen retrieval buffer (100X citrate buffer pH 6.0) (ab94674). Samples were incubated with the primary antibody (1/100) for 14 hours at 4°C. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG polyclonal (1/300) was used as the secondary antibody.

    See Abreview

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (ab51067)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (ab51067)

    Immunohistochemical staining of paraffin embedded human clear cell kidney carcinoma with purified ab51067 at a working dilution of 1/100. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counterstained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0.

    PBS was used instead of the primary antibody as the negative control (inset).

  • Western blot - Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (ab51067)
    Western blot - Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (ab51067)
    All lanes : Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (ab51067) at 1/1000 dilution

    Lane 1 : Mouse thymus tissue lysate
    Lane 2 : Rat thymus tissue lysate
    Lane 3 : Mouse lung tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 156 kDa
    Observed band size: 240 kDa why is the actual band size different from the predicted?



    Lanes 1 - 3: Merged signal (red and green). Green - ab51067 observed at 240 kDa. Red - loading control, ab18058, observed at 130 kDa.

    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab51067 and ab18058 (loading control) overnight at 4°C. Antibody binding was detected using Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at a 1:10000 dilution for 1hr at room temperature and then imaged.

  • ELISA - Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (ab51067)
    ELISA - Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (ab51067)

    ELISA analysis of Human Met recombinant protein at 1000 ng/mL with ab51067. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/2500 dilution was used as the secondary antibody.

  • Western blot - Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (ab51067)
    Western blot - Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (ab51067)
    All lanes : Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (ab51067) at 1/2000 dilution (purified)

    Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate
    Lane 2 : HEK-293 (Human epithelial cell line from embryonic kidney) cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution

    Predicted band size: 156 kDa
    Observed band size: 190 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunocytochemistry/ Immunofluorescence - Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (ab51067)
    Immunocytochemistry/ Immunofluorescence - Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (ab51067)

    Immunofluorescence staining of Jurkat (Human T cell leukemia cell line from peripheral blood) cells with purified ab51067 at a working dilution of 1/100, counterstained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel.

    The cells were fixed in 100% methanol and permeabilized using 0.1% Triton X 100.

    The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab51067 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.

  • Western blot - Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (ab51067)
    Western blot - Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (ab51067)

    Lane 1: Wild-type HAP1 cell lysate (40 µg)
    Lane 2: Met (c-Met) knockout HAP1 cell lysate (40 µg)
    Lane 3: HepG2 cell lysate (40µg) (40 µg)
    Lane 4: HEK-293 cel lysate (40µg) (40 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab51067 observed at 240 kDa. Red - loading control, ab18058, observed at 124 kDa.

    ab51067 was shown to specifically recognize Met (c-Met) in wild-type HAP1 cells along with additional cross reactive bands. No bands were observed when Met (c-Met) knockout samples were used. Wild-type and Met (c-Met) knockout samples were subjected to SDS-PAGE. ab51067 and ab18058 (loading control to Vinculin) were diluted at 1/1000 and 1/10,000 dilution respectively and incubated overnight at 4C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1hr at room temperature before imaging.

  • Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (ab51067)
    Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (ab51067)

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