Name: Anti-FADD antibody [EPR5030]
Brand: RabMAb
SKU: ab124812
Rating: 4 (9 reviews)

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Immunocytochemistry - 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

Carrier Free PE

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.

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

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
Recombinant Protein
- Recombinant Human FADD protein (ab89242)

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.

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

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)

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)

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) 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)

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)

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)

Flow cytometry protocols
Immunohistochemistry protocols
Western blot protocols

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

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

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1-7 of 7 Abreviews or Q&A

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

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

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

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

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

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

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

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

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

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

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

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

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

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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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Positive Controls

Recombinant Protein

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Sequence similarities

Domain

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

References (13)

Customer reviews and Q&As

Western blot abreview for Anti-FADD antibody [EPR5030]

Western blot abreview for Anti-FADD antibody [EPR5030]

Western blot abreview for Anti-FADD antibody [EPR5030]

Western blot abreview for Anti-FADD antibody [EPR5030]

Western blot abreview for Anti-FADD antibody [EPR5030]

Western blot abreview for Anti-FADD antibody [EPR5030]

Western blot abreview for Anti-FADD antibody [EPR5030]