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    fadd-antibody-epr5030-ab124812.pdf

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Cell Biology Apoptosis Intracellular Associated Proteins
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RecombinantRabMAb

Recombinant Anti-FADD antibody [EPR5030] (ab124812)

  • Datasheet
  • SDS
Reviews (9) Submit a question References (13)

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Immunocytochemistry - Anti-FADD antibody [EPR5030] (ab124812)
  • Western blot - Anti-FADD antibody [EPR5030] (ab124812)
  • Immunoprecipitation - Anti-FADD antibody [EPR5030] (ab124812)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FADD antibody [EPR5030] (ab124812)
  • Flow Cytometry (Intracellular) - Anti-FADD antibody [EPR5030] (ab124812)
  • Immunoprecipitation - Anti-FADD antibody [EPR5030] (ab124812)
  • Anti-FADD antibody [EPR5030] (ab124812)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR5030] to FADD
  • Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC, IP
  • Reacts with: Mouse

Conjugates logo Related conjugates and formulations

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Overview

  • Product name

    Anti-FADD antibody [EPR5030]
    See all FADD primary antibodies
  • Description

    Rabbit monoclonal [EPR5030] to FADD
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC, IPmore details
  • Species reactivity

    Reacts with: Mouse
  • Immunogen

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

  • Positive control

    • IHC-P: Mouse kidney tissue; ICC: NIH/3T3 cells; Flow Cyt (intra): NIH/3T3 cells. WB: NIH/3T3 whole cell lysate. IP: NIH/3T3 whole cell lysate and RAW264.7 whole cell lysate.
  • 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.

    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.

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

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 long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 0.05% BSA, 40% Glycerol (glycerin, glycerine), 59% PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR5030
  • 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

    • PE Anti-FADD antibody [EPR5030] (ab211700)
    • Anti-FADD antibody [EPR5030] - BSA and Azide free (ab229444)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • NIH/3T3 whole cell lysate (ab7179)
    • RAW 264.7 whole cell lysate (ab7187)
    • Mouse spleen tissue lysate - total protein (ab7937)
  • Recombinant Protein

    • Recombinant Human FADD protein (ab89242)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab124812 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
Flow Cyt (Intra)
1/40.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.  

WB (7)
1/1000 - 1/10000. Detects a band of approximately 28 kDa (predicted molecular weight: 23 kDa).
IHC-P
1/50 - 1/150. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

ICC
1/50.
IP (1)
1/30.
Notes
Flow Cyt (Intra)
1/40.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.  

WB
1/1000 - 1/10000. Detects a band of approximately 28 kDa (predicted molecular weight: 23 kDa).
IHC-P
1/50 - 1/150. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

ICC
1/50.
IP
1/30.

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: 14082 Mouse
    • SwissProt: Q61160 Mouse
    • Unigene: 5126 Mouse
    • 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

    • Immunocytochemistry - Anti-FADD antibody [EPR5030] (ab124812)
      Immunocytochemistry - Anti-FADD antibody [EPR5030] (ab124812)

      Immunocytochemistry analysis of NIH/3T3 (Mouse embryonic fibroblast) cells labeling FADD with purified ab124812 at 1/50 dilution (7.8 µg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

    • Western blot - Anti-FADD antibody [EPR5030] (ab124812)
      Western blot - Anti-FADD antibody [EPR5030] (ab124812)
      Anti-FADD antibody [EPR5030] (ab124812) at 1/1000 dilution (Purified) + NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 23 kDa

    • Immunoprecipitation - Anti-FADD antibody [EPR5030] (ab124812)
      Immunoprecipitation - Anti-FADD antibody [EPR5030] (ab124812)

      FADD was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 10 μg with ab124812 at 1/30 dilution (2 µg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab124812 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

      Lane 1: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 10 μg.

      Lane 2: NIH/3T3 whole cell lysate.

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab124812 in NIH/3T3 whole cell lysate.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Observed MW : 28 kDa.

      Exposure time: 41 secs.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FADD antibody [EPR5030] (ab124812)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FADD antibody [EPR5030] (ab124812)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue sections labeling FADD with purified ab124812 at 1/150 dilution (2.59 µg/mL). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1/0 dilution. PBS instead of the primary antibody was used as the negative control.
    • Flow Cytometry (Intracellular) - Anti-FADD antibody [EPR5030] (ab124812)
      Flow Cytometry (Intracellular) - Anti-FADD antibody [EPR5030] (ab124812)

      Intracellular Flow Cytometry analysis of NIH/3T3 (Mouse embryonic fibroblast) cells labelling FADD with purified ab124812 at 1/40 dilution (10 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).

       

    • Immunoprecipitation - Anti-FADD antibody [EPR5030] (ab124812)
      Immunoprecipitation - Anti-FADD antibody [EPR5030] (ab124812)

      FADD was immunoprecipitated from 0.35 mg RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate 10 μg with ab124812 at 1/30 dilution (2 µg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab124812 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

      Lane 1: RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate 10 μg.

      Lane 2: RAW264.7 whole cell lysate.

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab124812 in RAW264.7 whole cell lysate.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Observed MW : 28 kDa.

      Exposure time: 3 minutes.

    • Anti-FADD antibody [EPR5030] (ab124812)
      Anti-FADD antibody [EPR5030] (ab124812)

    Protocols

    • Flow cytometry protocols
    • Immunohistochemistry protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (13)

    Publishing research using ab124812? Please let us know so that we can cite the reference in this datasheet.

    ab124812 has been referenced in 13 publications.

    • Cai F  et al. ADT-OH, a hydrogen sulfide-releasing donor, induces apoptosis and inhibits the development of melanoma in vivo by upregulating FADD. Cell Death Dis 11:33 (2020). PubMed: 31949127
    • Liu H  et al. Copper Induces Oxidative Stress and Apoptosis in the Mouse Liver. Oxid Med Cell Longev 2020:1359164 (2020). PubMed: 32411316
    • Tummers B  et al. Caspase-8-Dependent Inflammatory Responses Are Controlled by Its Adaptor, FADD, and Necroptosis. Immunity 52:994-1006.e8 (2020). PubMed: 32428502
    • Li X  et al. Ubiquitination of RIPK1 regulates its activation mediated by TNFR1 and TLRs signaling in distinct manners. Nat Commun 11:6364 (2020). PubMed: 33311474
    • Zhang H  et al. Crucial Roles of the RIP Homotypic Interaction Motifs of RIPK3 in RIPK1-Dependent Cell Death and Lymphoproliferative Disease. Cell Rep 31:107650 (2020). PubMed: 32433959
    View all Publications for this product

    Customer reviews and Q&As

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

    1-7 of 7 Abreviews or Q&A

    Western blot abreview for Anti-FADD antibody [EPR5030]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Rabbit Cell lysate - whole cell (Rabbit Reticulocyte)
    Gel Running Conditions
    Reduced Denaturing (4-12% Bolt Bis-Tris Plus gel)
    Loading amount
    0.5 µg
    Specification
    Rabbit Reticulocyte
    Blocking step
    Li-Cor Odyssey Blocking Buffer (TBS) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Feb 09 2017

    Western blot abreview for Anti-FADD antibody [EPR5030]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Hamster Cell lysate - whole cell (BHK-21 (Kidney Fibroblasts))
    Gel Running Conditions
    Reduced Denaturing (4-12% Bolt Bis-Tris Plus gel (MOPS buffer))
    Loading amount
    1e+006 cells
    Specification
    BHK-21 (Kidney Fibroblasts)
    Blocking step
    Li-Cor Odyssey Blocking Buffer (TBS) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Feb 08 2017

    Western blot abreview for Anti-FADD antibody [EPR5030]

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Chinese Hamster Cell lysate - whole cell (Chinese hamster ovary (CHO) cells)
    Gel Running Conditions
    Reduced Denaturing (4-12% Bolt Bis-Tris Plus gel (MES buffer))
    Loading amount
    50000 cells
    Specification
    Chinese hamster ovary (CHO) cells
    Blocking step
    Li-Cor Odyssey Blocking Buffer (TBS) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Dec 12 2016

    Western blot abreview for Anti-FADD antibody [EPR5030]

    Inconclusive
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Dog Cell lysate - whole cell (Canine Kidney Epithelial MDCK cells)
    Gel Running Conditions
    Reduced Denaturing (4-12% Bolt Bis-Tris Plus gel)
    Loading amount
    500000 cells
    Specification
    Canine Kidney Epithelial MDCK cells
    Blocking step
    Li-Cor Odyssey Blocking Buffer (TBS) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Nov 11 2016

    Western blot abreview for Anti-FADD antibody [EPR5030]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Human Foreskin Fibroblasts & Mouse embryonic fibro)
    Gel Running Conditions
    Reduced Denaturing (4-12% Bolt Bis-Tris Plus gradient gel)
    Loading amount
    200000 cells
    Specification
    Human Foreskin Fibroblasts & Mouse embryonic fibro
    Blocking step
    Li-Cor Odyssey Blocking Buffer (TBS) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Feb 05 2016

    Western blot abreview for Anti-FADD antibody [EPR5030]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (mouse embryonic fibroblasts)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    10000 cells
    Treatment
    With or without 100uM zVAD
    Specification
    mouse embryonic fibroblasts
    Blocking step
    Li-Cor Odyssey Blocking Buffer (TBS) as blocking agent for 14 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 4°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    DR. Yi-Chieh Perng

    Verified customer

    Submitted Nov 30 2015

    Western blot abreview for Anti-FADD antibody [EPR5030]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    5 µg
    Gel Running Conditions
    Reduced Denaturing (4-12)
    Sample
    Mouse Tissue lysate - whole (uterus)
    Specification
    uterus
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Dr. Varkha Agrawal

    Verified customer

    Submitted Jan 07 2014

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