Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Axl antibody [EPR23892-15] (ab259831)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23892-15] to Axl
  • Suitable for: ICC, Flow Cyt, WB
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-Axl antibody [EPR23892-15]
    See all Axl primary antibodies

  • Description

    Rabbit monoclonal [EPR23892-15] to Axl

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Wild-type HeLa whole cell lysate; NCI-H1299 whole cell lysate; DU145 whole cell lysate. ICC: NCI-H1299; Hela cells. Flow Cyt: Hela cells; NCI-H1299 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.

Properties

Images

  • Western blot - Anti-Axl antibody [EPR23892-15] (ab259831)
    Western blot - Anti-Axl antibody [EPR23892-15] (ab259831)
    All lanes : Anti-Axl antibody [EPR23892-15] (ab259831) at 1/1000 dilution

    Lane 1 : Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : Axl knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 3 : NCI-H1299 (human lung carcinoma epithelial cell) whole cell lysate
    Lane 4 : Jurkat (human T cell leukemia T lymphocyte), whole cell lysate

    Lysates/proteins at 40 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (ab216773) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (ab216776) at 1/10000 dilution

    Predicted band size: 98 kDa
    Observed band size: 140,80 kDa
    why is the actual band size different from the predicted?



    Blocking and diluting buffer and concentration: 5% NFDM/TBST

    Lanes 1-4: Merged signal (red and green). Green - ab259831 observed at 140, 80 kDa. Red - loading control ab8245 observed at 36 kDa.

    ab259831 Anti-Axl antibody [EPR23892-15] was shown to react with Axl in Hela cells in Western blot. Loss of signal was observed when knockout cell line ab261810 (knockout cell lysate ab257151) was used. Wild-type and Axl knockout samples were subjected to SDS-PAGE.

    ab259831 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at 4°C overnight 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 10000 dilution for 1 hour at room temperature before imaging.

    Negative control: Jurkat (PMID: 28423548).

     

  • Immunocytochemistry - Anti-Axl antibody [EPR23892-15] (ab259831)
    Immunocytochemistry - Anti-Axl antibody [EPR23892-15] (ab259831)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Trito nX-100 permeabilized NCI-H1299 cells labelling Axl with ab259831 at 1/500 (0.98 µg/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing strong membranous and weak cytoplasmic staining in NCI-H1299 cells (PMID: 27443522). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).

    Negative control: Jurkat (PMID: 28423548).

    Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

  • Flow Cytometry - Anti-Axl antibody [EPR23892-15] (ab259831)
    Flow Cytometry - Anti-Axl antibody [EPR23892-15] (ab259831)

    Flow cytometric analysis of Parental HeLa (human cervix adenocarcinoma epithelial cell (Right panel)/ AXL knockout HeLa cells (Left panel) cells labelling Axl with ab259831 at 1/500 dilution (0.1µg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

    Positive staining on HeLa cells (ab255928), while no staining on AXL knockout HeLa cells (ab261810).

    Gated on viable cells.

  • Western blot - Anti-Axl antibody [EPR23892-15] (ab259831)
    Western blot - Anti-Axl antibody [EPR23892-15] (ab259831)
    All lanes : Anti-Axl antibody [EPR23892-15] (ab259831) at 1/1000 dilution

    Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
    Lane 2 : DU145 (human prostate carcinoma epithelial cell), whole cell lysate
    Lane 3 : NCI-H1299 (human lung carcinoma epithelial cell), whole cell lysate
    Lane 4 : Jurkat (human T cell leukemia T lymphocyte), whole cell lysate

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size: 98 kDa
    Observed band size: 140,80 kDa why is the actual band size different from the predicted?



    Blocking and diluting buffer and concentration: 5% NFDM/TBST

    Negative control: Jurkat (PMID: 28423548).

    Exposure time: 37 seconds

     

  • Immunocytochemistry - Anti-Axl antibody [EPR23892-15] (ab259831)
    Immunocytochemistry - Anti-Axl antibody [EPR23892-15] (ab259831)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Axl KO HeLa (ab261810) cells labelling Axl with ab259831 at 1/100 (4.9 µg/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing strong membranous and weak cytoplasmic staining in parental HeLa cells (ab255928), while no staining in AXL knockout HeLa cells (ab261810). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

  • Flow Cytometry - Anti-Axl antibody [EPR23892-15] (ab259831)
    Flow Cytometry - Anti-Axl antibody [EPR23892-15] (ab259831)

    Flow cytometric analysis of Jurkat (Human T cell leukemia T lymphocyte, Left panel)/ NCI-H1299 (human lung carcinoma epithelial cell, Right panel) cells labelling Axl with ab259831 at 1/500 dilution (0.1µg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

    Negative control: Jurkat (PMID: 28423548).

    Gated on viable cells.

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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