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, IP, ICC/IF
- 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, IP, ICC/IFmore 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/IF: 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.
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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IP | (1) |
1/30.
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ICC/IF |
1/50.
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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. |
IP
1/30. |
ICC/IF
1/50. |
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) 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 (23)
ab124812 has been referenced in 23 publications.
- Li X et al. Caspase-8 auto-cleavage regulates programmed cell death and collaborates with RIPK3/MLKL to prevent lymphopenia. Cell Death Differ 29:1500-1512 (2022). PubMed: 35064213
- Parshenkov A & Hennet T Glycosylation-Dependent Induction of Programmed Cell Death in Murine Adenocarcinoma Cells. Front Immunol 13:797759 (2022). PubMed: 35222379
- Li M et al. Ubiquitin-binding domain in ABIN1 is critical for regulating cell death and inflammation during development. Cell Death Differ 29:2034-2045 (2022). PubMed: 35430614
- Yu J et al. Effects and potential mechanism of Ca2+/calmodulin‑dependent protein kinase II pathway inhibitor KN93 on the development of ovarian follicle. Int J Mol Med 50:N/A (2022). PubMed: 35929517
- Wu G et al. PP6 negatively modulates LUBAC-mediated M1-ubiquitination of RIPK1 and c-FLIPL to promote TNFα-mediated cell death. Cell Death Dis 13:773 (2022). PubMed: 36071040