Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Fas antibody [EPR5700] (ab133619)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR5700] to Fas
  • Suitable for: WB, IHC-P, ICC
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-Fas antibody [EPR5700]
    See all Fas primary antibodies

  • Description

    Rabbit monoclonal [EPR5700] to Fas

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    ICC
    Human
    IHC-P
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human Fas aa 150 to the C-terminus. The exact sequence is proprietary.
    Database link: P25445

  • Positive control

    • WB: HeLa, Ramos, HT-1080, and Raji cell lysates. IHC-P: Human tonsil tissue. ICC: Raji cells

  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

     

    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-Fas antibody [EPR5700] (ab133619)
    Western blot - Anti-Fas antibody [EPR5700] (ab133619)
    All lanes : Anti-Fas antibody [EPR5700] (ab133619) at 1/1000 dilution

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

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 37 kDa
    Observed band size: 37 kDa



    Lanes 1-2: Merged signal (red and green). Green - ab133619 observed at 37 kDa. Red - loading control ab8245 observed at 37 kDa.

     ab133619 Anti-Fas antibody [EPR5700] was shown to specifically react with Fas in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265260 (knockout cell lysate ab256911) was used. Wild-type and Fas knockout samples were subjected to SDS-PAGE. ab133619 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 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.

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fas antibody [EPR5700] (ab133619)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fas antibody [EPR5700] (ab133619)

    Anti-Fas antibody [EPR5700] (ab133619)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling Fas with ab133619 at a dilution of 1:500. Heat mediated antigen retrieval was performed using AR9 antigen retrieval solution, and microwave treatment for 15 min at 20% power. Anti-Rabbit/Mouse HRP polymer (PerkinElmer Opal Polymer HRP Ms Plus Rb) was used as secondary antibody. Opal tyramide amplification was performed using Opal 520 fluorophore. Counterstained with DAPI stain. Image scanned with Vectra 3.0 and analyzed via Phenochart software.
    This image was courteously provided by Dr. Houssein Abdul Sater, Georgia Cancer Center.

  • Immunocytochemistry - Anti-Fas antibody [EPR5700] (ab133619)
    Immunocytochemistry - Anti-Fas antibody [EPR5700] (ab133619)
    Immunocytochemistry analysis of Raji (Human Burkitt's lymphoma B lymphocyte) labeling Fas with purified ab133619 at 1/50 dilution. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/1000 (2 µg/ml) was used as the secondary antibody. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.31 µg/ml) was used as counterstain. Nuclei were stained blue with DAPI.
    Negative control: PBS instead of the primary antibody.
  • Western blot - Anti-Fas antibody [EPR5700] (ab133619)
    Western blot - Anti-Fas antibody [EPR5700] (ab133619)
    All lanes : Anti-Fas antibody [EPR5700] (ab133619) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 cell lysate
    Lane 2 : Fas knockout HAP1 cell lysate
    Lane 3 : Raji cell lysate
    Lane 4 : Ramos cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 37 kDa



    Lanes 1 - 4: Merged signal (red and green). Green - ab133619 observed at 42 kDa. Red - loading control, ab8245, observed at 38 kDa.

    ab133619 was shown to specifically react with Fas when Fas knockout samples were used. Wild-type and Fas knockout samples were subjected to SDS-PAGE. ab133619 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10 000 and incubated overnight at 4°C. 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 1 h at room temperature before imaging.

  • Western blot - Anti-Fas antibody [EPR5700] (ab133619)
    Western blot - Anti-Fas antibody [EPR5700] (ab133619)
    All lanes : Anti-Fas antibody [EPR5700] (ab133619) at 1/1000 dilution

    Lane 1 : Ramos cell lysate
    Lane 2 : HT-1080 cell lysate
    Lane 3 : Raji cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

    Predicted band size: 37 kDa



    Actual band size : 45 kDa
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fas antibody [EPR5700] (ab133619)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fas antibody [EPR5700] (ab133619)

    Immunohistochemical analysis of paraffin embedded Human tonsil tissue labelling CD95 with ab133619 antibody at a dilution of 1/250.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Western blot - Anti-Fas antibody [EPR5700] (ab133619)
    Western blot - Anti-Fas antibody [EPR5700] (ab133619)
    All lanes : Anti-Fas antibody [EPR5700] (ab133619)

    Lane 1 : Wild-type HAP1 cell lysate
    Lane 2 : Fas knockout HAP1 cell lysate
    Lane 3 : Raji cell lysate
    Lane 4 : Ramos cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 37 kDa



    Lanes 1 - 4: Merged signal (red and green). Green - ab133619 observed at 42 kDa. Red - loading control, ab8245, observed at 38 kDa.

    This western blot image is a comparison between ab133619 and a competitor's top cited rabbit polyclonal antibody.

  • Anti-Fas antibody [EPR5700] (ab133619)
    Anti-Fas antibody [EPR5700] (ab133619)

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