Recombinant 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
Related conjugates and formulations
Overview
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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.
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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.
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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
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
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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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR5030 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
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 |
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Flow Cyt (Intra) |
1/40.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB | (7) |
1/1000 - 1/10000. Detects a band of approximately 28 kDa (predicted molecular weight: 23 kDa).
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IHC-P |
1/50 - 1/150. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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ICC |
1/50.
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IP | (1) |
1/30.
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Notes |
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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. |
ICC
1/50. |
IP
1/30. |
Target
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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. - Information by UniProt
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Database links
- Entrez Gene: 14082 Mouse
- SwissProt: Q61160 Mouse
- Unigene: 5126 Mouse
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Alternative names
- FADD antibody
- FADD protein antibody
- FADD_HUMAN antibody
see all
Images
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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.
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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 -
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.
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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.
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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).
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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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (13)
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