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Cell Biology Apoptosis Receptors Death Receptors
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Fas antibody [EPR5700] - BSA and Azide free (ab178076)

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Western blot - Anti-Fas antibody [EPR5700] - BSA and Azide free (ab178076)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fas antibody [EPR5700] - BSA and Azide free (ab178076)
  • Immunocytochemistry/ Immunofluorescence - Anti-Fas antibody [EPR5700] - BSA and Azide free (ab178076)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fas antibody [EPR5700] - BSA and Azide free (ab178076)
  • Immunocytochemistry/ Immunofluorescence - Anti-Fas antibody [EPR5700] - BSA and Azide free (ab178076)
  • Immunocytochemistry/ Immunofluorescence - Anti-Fas antibody [EPR5700] - BSA and Azide free (ab178076)
  • Anti-Fas antibody [EPR5700] - BSA and Azide free (ab178076)

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 - BSA and Azide free
  • Suitable for: WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 Unconjugated

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Knockout
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Human FAS knockout HeLa cell line (ab265260)

View more associated products

Overview

  • Product name

    Anti-Fas antibody [EPR5700] - BSA and Azide free
    See all Fas primary antibodies
  • Description

    Rabbit monoclonal [EPR5700] to Fas - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IFmore details
    Unsuitable for: Flow Cyt (Intra) or IP
  • Species reactivity

    Reacts with: Human
  • Immunogen

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

  • Positive control

    • WB: HeLa cell lysate. IHC-P: Human tonsil tissue. ICC/IF: Raji cells
  • General notes

    ab178076 is the carrier-free version of ab133619.

    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 <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C. Do Not Freeze.
  • Storage buffer

    Constituent: PBS
  • Carrier free

    Yes
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR5700
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Apoptosis
    • Receptors
    • Death Receptors
    • Immunology
    • Cell Type Markers
    • CD
    • Cytokines
    • Immunology
    • Innate Immunity
    • Cytokines
    • TNF Superfamily

Associated products

  • Alternative Versions

    • Anti-Fas antibody [EPR5700] (ab133619)
    • Alexa Fluor® 488 Anti-Fas antibody [EPR5700] (ab204670)
    • Alexa Fluor® 647 Anti-Fas antibody [EPR5700] (ab204671)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human FAS knockout HeLa cell line (ab265260)
  • KO cell lysates

    • Human FAS knockout HeLa cell lysate (ab256911)
  • Positive Controls

    • Raji membrane extract lysate (ab30126)
    • Ramos whole cell lysate (ab3955)
  • Recombinant Protein

    • Recombinant human Fas protein (ab155635)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab178076 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
Use at an assay dependent concentration. Detects a band of approximately 45 kDa (predicted molecular weight: 37 kDa).
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF
Use at an assay dependent concentration.
Notes
WB
Use at an assay dependent concentration. Detects a band of approximately 45 kDa (predicted molecular weight: 37 kDa).
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF
Use at an assay dependent concentration.
Application notes
Is unsuitable for Flow Cyt (Intra) or 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.
  • Target information above from: UniProt accession P25445 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 355 Human
    • Omim: 134637 Human
    • SwissProt: P25445 Human
    • Unigene: 244139 Human
    • 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] - BSA and Azide free (ab178076)
      Western blot - Anti-Fas antibody [EPR5700] - BSA and Azide free (ab178076)
      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



      This data was developed using the same antibody clone in a different buffer formulation (ab133619).

      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] - BSA and Azide free (ab178076)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fas antibody [EPR5700] - BSA and Azide free (ab178076)

      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/ Immunofluorescence - Anti-Fas antibody [EPR5700] - BSA and Azide free (ab178076)
      Immunocytochemistry/ Immunofluorescence - Anti-Fas antibody [EPR5700] - BSA and Azide free (ab178076)
      This data was developed using the same antibody clone in a different buffer formulation (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.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fas antibody [EPR5700] - BSA and Azide free (ab178076)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fas antibody [EPR5700] - BSA and Azide free (ab178076)

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

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133619).

      Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

    • Immunocytochemistry/ Immunofluorescence - Anti-Fas antibody [EPR5700] - BSA and Azide free (ab178076)
      Immunocytochemistry/ Immunofluorescence - Anti-Fas antibody [EPR5700] - BSA and Azide free (ab178076)

      Clone EPR5700 (ab178076) has been successfully conjugated by Abcam. This image was generated using Anti-Fas antibody [EPR5700] (Alexa Fluor® 488). Please refer to ab204670 for protocol details.

      ab204670 staining Fas in Raji cells. The cells were fixed with 80% methanol (5 min) and then incubated in 1%BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated overnight at +4°C with ab204670 at 1/100 dilution (shown in green) and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

      Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

      This product also gave a positive signal under the same testing conditions in Raji cells fixed with 4% formaldehyde (10 min).

    • Immunocytochemistry/ Immunofluorescence - Anti-Fas antibody [EPR5700] - BSA and Azide free (ab178076)
      Immunocytochemistry/ Immunofluorescence - Anti-Fas antibody [EPR5700] - BSA and Azide free (ab178076)

      Clone EPR5700 (ab178076) has been successfully conjugated by Abcam. This image was generated using Anti-Fas antibody [EPR5700] (Alexa Fluor® 647). Please refer to ab204671 for protocol details.

      ab204671 staining Fas in Raji cells. The cells were fixed with 4% formaldehyde (10 min) and then incubated in 1%BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated overnight at +4°C with ab204671 at 1/50 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 488), at 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

       

      Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    • Anti-Fas antibody [EPR5700] - BSA and Azide free (ab178076)
      Anti-Fas antibody [EPR5700] - BSA and Azide free (ab178076)

    Protocols

    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    References (0)

    Publishing research using ab178076? Please let us know so that we can cite the reference in this datasheet.

    ab178076 has not yet been referenced specifically in any publications.

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