Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-Fas antibody [EPR5700] (ab133619)

Submit a review Q&A (1)References (15)

Overview

  • Product name

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

  • Description

    Rabbit monoclonal [EPR5700] to Fas

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IFmore details
    Unsuitable for: IP

  • Species reactivity

    Reacts with: Human

  • 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.

  • General notes

    A trial size is available to purchase for this antibody.

    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

Applications

Our Abpromise guarantee covers the use of ab133619 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
WB 1/1000 - 1/10000. Detects a band of approximately 45 kDa (predicted molecular weight: 37 kDa).
IHC-P 1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF 1/250 - 1/500.
  • Application notes
    Is unsuitable for IP.

    • Target

    • Function

      Receptor for TNFSF6/FASLG. The adapter molecule FADD recruits caspase-8 to the activated receptor. The resulting death-inducing signaling complex (DISC) performs caspase-8 proteolytic activation which initiates the subsequent cascade of caspases (aspartate-specific cysteine proteases) mediating apoptosis. FAS-mediated apoptosis may have a role in the induction of peripheral tolerance, in the antigen-stimulated suicide of mature T-cells, or both. The secreted isoforms 2 to 6 block apoptosis (in vitro).

    • Tissue specificity

      Isoform 1 and isoform 6 are expressed at equal levels in resting peripheral blood mononuclear cells. After activation there is an increase in isoform 1 and decrease in the levels of isoform 6.

    • Involvement in disease

      Defects in FAS are the cause of autoimmune lymphoproliferative syndrome type 1A (ALPS1A) [MIM:601859]; also known as Canale-Smith syndrome (CSS). ALPS is a childhood syndrome involving hemolytic anemia and thrombocytopenia with massive lymphadenopathy and splenomegaly.

    • Sequence similarities

      Contains 1 death domain.
      Contains 3 TNFR-Cys repeats.

    • Domain

      Contains a death domain involved in the binding of FADD, and maybe to other cytosolic adapter proteins.

    • Cellular localization

      Secreted and Cell membrane.
    • Information by UniProt

    • Database links

      • Alternative names

        • ALPS 1A antibody
        • ALPS1A antibody
        • APO 1 antibody
        • Apo 1 antigen antibody
        • APO 1 cell surface antigen antibody
        • Apo-1 antigen antibody
        • APO1 antibody
        • Apo1 antigen antibody
        • APO1 cell surface antigen antibody
        • Apoptosis antigen 1 antibody
        • Apoptosis mediating surface antigen FAS antibody
        • Apoptosis-mediating surface antigen FAS antibody
        • APT 1 antibody
        • APT1 antibody
        • CD 95 antibody
        • CD 95 antigen antibody
        • CD95 antibody
        • CD95 antigen antibody
        • Delta Fas antibody
        • Delta Fas/APO 1/CD95 antibody
        • Delta Fas/APO1/CD95 antibody
        • Fas (TNF receptor superfamily, member 6) antibody
        • FAS 1 antibody
        • FAS 827dupA antibody
        • Fas AMA antibody
        • Fas antibody
        • FAS Antigen antibody
        • Fas cell surface death receptor antibody
        • FAS1 antibody
        • FASLG receptor antibody
        • FASTM antibody
        • sFAS antibody
        • Surface antigen APO1 antibody
        • TNF receptor superfamily, member 6 antibody
        • TNFRSF 6 antibody
        • TNFRSF6 antibody
        • TNR6_HUMAN antibody
        • Tumor necrosis factor receptor superfamily member 6 antibody
        see all

      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 - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

        ab133619 was shown to react with Fas in wild-type HeLa cells in western blot. Loss of signal was observed when knockout sample ab256911 was used. Wild-type HeLa and FAS knockout HeLa cell lysates were subjected to SDS-PAGE. ab133619 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 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.

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

        Lane 1: Wild-type HAP1 cell lysate (20 µg)
        Lane 2: Fas knockout HAP1 cell lysate (20 µg)
        Lane 3: Raji cell lysate (20 µg)
        Lane 4: Ramos cell lysate (20 µg)
        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)

        Lane 1: Wild-type HAP1 cell lysate (20 µg)
        Lane 2: Fas knockout HAP1 cell lysate (20 µg)
        Lane 3: Raji cell lysate (20 µg)
        Lane 4: Ramos cell lysate (20 µg)
        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.

      References

      This product has been referenced in:

      • Su X  et al. Mechanical, nanomorphological and biological reconstruction of early-stage apoptosis in HeLa cells induced by cytochalasin B. Oncol Rep 41:928-938 (2019). Read more (PubMed: 30535459) »
      • Zhang L  et al. Cysteine-rich intestinal protein 1 suppresses apoptosis and chemosensitivity to 5-fluorouracil in colorectal cancer through ubiquitin-mediated Fas degradation. J Exp Clin Cancer Res 38:120 (2019). Read more (PubMed: 30850009) »
      See all 15 Publications for this product

      Publishing research using ab133619? Please let us know so that we can cite the reference in this datasheet.

      Customer reviews and Q&As

      Show All Reviews Q&A
      Submit a review Submit a question

      Question

      SEARCHING ANTIBODY FOR FLOW CYTOMETRY AGAINST HUMAN CELLS BUT NOT MOUSE CELLS

      Dear,
      Parhaps ab133619 will also work. This CD95, it is the same as Fas, right? If so, this should be expressed on the outer membrane by PC3 cells, according to literature, so that may be a safer one than the cytokeratin.
      Best,
      Thank you very much for this info. ab32118 may be a good candidate.
      I have one major question though, which I cannot really find in literature. I am using PC3 cells as human cancer cells. They are reported to express cytokeratin 18. However, is this protein present at the cell surface in living cells? I find that in most studies cells are fixed with for example aceton, before the staining of keratins. So when you tested this antibody in flow cyt, on which cells was this tested, and were those cells fixed/permeabilized in some way or were they alive?
      Thank you.

      Read More
      Answer

      Thank you very much for your answer.
      I have contacted the laboratory to gather some more information in regards to the species testing of these antibodies. I am sorry it has taken so long time. Have you already proceeded meanwhile?
      In regards to the cytokeratin 18 antibody - as far as I know, the cytokeratins are all cytoplasmic. The ab32118 has been tested in flow cytometry on permeabilized cells (with methanol).
      In regards to the CD95 antibody ab133619, we do not think it crossreacts with mouse and rat as the immunogen shares only 45% identity with mouse and rat CD95. However there was a signal in mouse heart for which we could not determine whether it was specific or not. I am very sorry for not being able to provide you with a better answer still. What I however would like to offer is a testing discount: you would buy this antibody and test it. You would then provide the feedback via an Abreview. Once we have the feedback, you would receive an antibody of your choice from our catalog for free.
      If such a testing would interest you, please do let me know and I will describe you the offer in more details.
      If however you prefer looking at other antibodies, please let me know as well. I would be pleased to investigate also other antibodies of the list for you (and I would expect it being faster this time .. ).
      Please let me know if you have any question. I look forward to hear back from you.

      Read More

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