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    fadd-antibody-epr4415-ab108601.pdf

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Cell Biology Apoptosis Intracellular Associated Proteins
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-FADD antibody [EPR4415] (ab108601)

  • Datasheet
  • SDS
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Western blot - Anti-FADD antibody [EPR4415] (ab108601)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FADD antibody [EPR4415] (ab108601)
  • Flow Cytometry - Anti-FADD antibody [EPR4415] (ab108601)
  • Western blot - Anti-FADD antibody [EPR4415] (ab108601)
  • Western blot - Anti-FADD antibody [EPR4415] (ab108601)
  • Anti-FADD antibody [EPR4415] (ab108601)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR4415] to FADD
  • Suitable for: WB, IP, IHC-P, Flow Cyt
  • Knockout validated
  • Reacts with: Human

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Knockout
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Human FADD knockout HeLa cell line (ab261817)

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Overview

  • Product name

    Anti-FADD antibody [EPR4415]
    See all FADD primary antibodies
  • Description

    Rabbit monoclonal [EPR4415] to FADD
  • Host species

    Rabbit
  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    IHC-P
    Human
    WB
    Human
    See all applications and species data
  • Immunogen

    Synthetic peptide within Human FADD aa 1-150. The exact sequence is proprietary.
    Database link: Q13158

  • Positive control

    • WB: A431, Jurkat, HeLa, and SKBR-3 cell lysates. IHC-P: Human kidney tissue. Flow Cyt: A431 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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.20
    Preservative: 0.05% Sodium azide
    Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Tissue culture supernatant
  • Clonality

    Monoclonal
  • Clone number

    EPR4415
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Apoptosis
    • Intracellular
    • Associated Proteins
    • Cancer
    • Invasion/microenvironment
    • Apoptosis
    • Death receptors & ligands
    • FADD
    • Cancer
    • Cell Death
    • Apoptosis
    • Receptors
    • Death receptors & ligands
    • FADD
    • Cancer
    • Cell Death
    • Necroptosis

Associated products

  • Alternative Versions

    • Alexa Fluor® 647 Anti-FADD antibody [EPR4415] (ab207364)
    • Alexa Fluor® 594 Anti-FADD antibody [EPR4415] (ab211975)
    • Anti-FADD antibody [EPR4415] - BSA and Azide free (ab232045)
  • Compatible Secondaries

    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

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

    • Human FADD knockout HeLa cell line (ab261817)
  • KO cell lysates

    • Human FADD knockout HeLa cell lysate (ab257261)
  • KO cell pellets

    • Human FADD knockout HeLa cell pellet (ab279006)
  • Positive Controls

    • HeLa whole cell lysate (ab29545)
    • Jurkat whole cell lysate (ab7899)
    • A-431 whole cell lysate (ab7909)
  • Recombinant Protein

    • Recombinant Human FADD protein (ab89242)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab108601 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
Flow Cyt
Human
IHC-P
Human
WB
Human
Application Abreviews Notes
WB
1/1000 - 1/10000. Detects a band of approximately 28 kDa (predicted molecular weight: 23 kDa).
IP
1/10 - 1/100.
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Flow Cyt
Use at an assay dependent concentration.
Notes
WB
1/1000 - 1/10000. Detects a band of approximately 28 kDa (predicted molecular weight: 23 kDa).
IP
1/10 - 1/100.
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Flow Cyt
Use at an assay dependent concentration.

Target

  • Function

    Apoptotic adaptor molecule that recruits caspase-8 or caspase-10 to the activated Fas (CD95) or TNFR-1 receptors. The resulting aggregate called the death-inducing signaling complex (DISC) performs caspase-8 proteolytic activation. Active caspase-8 initiates the subsequent cascade of caspases mediating apoptosis.
  • Tissue specificity

    Expressed in a wide variety of tissues, except for peripheral blood mononuclear leukocytes.
  • Sequence similarities

    Contains 1 death domain.
    Contains 1 DED (death effector) domain.
  • Domain

    Contains a death domain involved in the binding of the corresponding domain within Fas receptor.
  • Target information above from: UniProt accession Q13158 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 8772 Human
    • Omim: 602457 Human
    • SwissProt: Q13158 Human
    • Unigene: 86131 Human
    • Alternative names

      • FADD antibody
      • FADD protein antibody
      • FADD_HUMAN antibody
      • Fas (TNFRSF6) associated via death domain antibody
      • Fas associated via death domain antibody
      • Fas associating death domain containing protein antibody
      • Fas associating protein antibody
      • Fas associating protein with death domain antibody
      • Fas TNFRSF6 associated via death domain antibody
      • FAS-associated death domain protein antibody
      • FAS-associating death domain-containing protein antibody
      • GIG 3 antibody
      • GIG3 antibody
      • Growth inhibiting gene 3 protein antibody
      • Growth-inhibiting gene 3 protein antibody
      • H sapiens mRNA for mediator of receptor induced toxicity antibody
      • Mediator of receptor induced toxicity antibody
      • MGC8528 antibody
      • MORT 1 antibody
      • MORT1 antibody
      • Protein FADD antibody
      see all

    Images

    • Western blot - Anti-FADD antibody [EPR4415] (ab108601)
      Western blot - Anti-FADD antibody [EPR4415] (ab108601)
      All lanes : Anti-FADD antibody [EPR4415] (ab108601) at 1/1000 dilution

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

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 23 kDa
      Observed band size: 23 kDa



      Lanes 1 - 2: Merged signal (red and green). Green - ab108601 observed at 23 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

      ab108601 was shown to react with FADD in wild-type HeLa cells in western blot with loss of signal observed in FADD knockout cell line ab261817 (FADD knockout cell lysate ab257261). Wild-type and FADD knockout HeLa cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab108601 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FADD antibody [EPR4415] (ab108601)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FADD antibody [EPR4415] (ab108601)

      Immunohistochemical staining of paraffin-embedded Human kidney tissue using ab108601 at a dilution of 1/100.

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

    • Flow Cytometry - Anti-FADD antibody [EPR4415] (ab108601)
      Flow Cytometry - Anti-FADD antibody [EPR4415] (ab108601)

      Flow Cytometry analysis of A431 (human epidermoid carcinoma) cells labeling FADD with purified ab108601 at 1/140 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488)(1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.

    • Western blot - Anti-FADD antibody [EPR4415] (ab108601)
      Western blot - Anti-FADD antibody [EPR4415] (ab108601)
      All lanes : Anti-FADD antibody [EPR4415] (ab108601) at 1/1000 dilution

      Lane 1 : Wild-type HAP1 cell lysate
      Lane 2 : FADD knockout HAP1 cell lysate
      Lane 3 : A431 cell lysate
      Lane 4 : Jurkat cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 23 kDa



      Lanes 1 - 4: Merged signal (red and green). Green - ab108601 observed at 25 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab108601 was shown to specifically react with FADD when FADD knockout samples were used. Wild-type and FADD knockout samples were subjected to SDS-PAGE. ab108601 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/2000 respectively 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-FADD antibody [EPR4415] (ab108601)
      Western blot - Anti-FADD antibody [EPR4415] (ab108601)
      All lanes : Anti-FADD antibody [EPR4415] (ab108601) at 1/1000 dilution

      Lane 1 : A431 cell lysate
      Lane 2 : Jurkat cell lysate
      Lane 3 : HeLa cell lysate
      Lane 4 : SKBR-3 cell lysate

      Lysates/proteins at 10 µg per lane.

      Predicted band size: 23 kDa

    • Anti-FADD antibody [EPR4415] (ab108601)
      Anti-FADD antibody [EPR4415] (ab108601)

    Protocols

    • Flow cytometry protocols
    • Immunoprecipitation protocols
    • Immunohistochemistry protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (5)

    Publishing research using ab108601? Please let us know so that we can cite the reference in this datasheet.

    ab108601 has been referenced in 5 publications.

    • Zeng L  et al. Prognostic value of biomarkers EpCAM and aB-crystallin associated with lymphatic metastasis in breast cancer by iTRAQ analysis. BMC Cancer 19:831 (2019). PubMed: 31443698
    • Ali M & Mocarski ES Proteasome inhibition blocks necroptosis by attenuating death complex aggregation. Cell Death Dis 9:346 (2018). PubMed: 29497034
    • Li X  et al. SPAG6 regulates cell apoptosis through the TRAIL signal pathway in myelodysplastic syndromes. Oncol Rep 37:2839-2846 (2017). PubMed: 28393201
    • Liu Y  et al. MiR-7a is an important mediator in Fas-associated protein with death domain (FADD)-regulated expression of focal adhesion kinase (FAK). Oncotarget 7:51393-51407 (2016). WB . PubMed: 27286445
    • Lu YM  et al. P2X7 signaling promotes microsphere embolism-triggered microglia activation by maintaining elevation of Fas ligand. J Neuroinflammation 9:172 (2012). WB ; Mouse . PubMed: 22789015

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