![Sandwich ELISA - Smad3 antibody [EP568Y] (ab40854) image](#Smad3-Primary-antibodies-ab40854-1.jpg)
![Western blot - Smad3 antibody [EP568Y] (ab40854) image](#ab40854_1.jpg)
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Smad3 antibody [EP568Y] (ab40854) image](#ab40854_2.jpg)
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Products:Signal Transduction >> Signaling Pathway >> Nuclear Signaling >> SMADs
Overview
Product name
Anti-Smad3 antibody [EP568Y]
See all Smad3 products (21) ...
Description
Rabbit monoclonal [EP568Y] to Smad3
Specificity
This antibody is specific for Smad3.
Tested applications
WB, ICC, IHC-P, IP, Sandwich ELISAmore details
Cross reactivity
Reacts with
Mouse, Rat, Human
Immunogen
Synthetic peptide corresponding to the internal region of Human Smad3.
Positive control
Jurkat cell lysate and human prostate carcinoma tissue.
General notes
Produced under U.S. Patent No. 5,675,063.
Properties
Form
Liquid
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage buffer
Preservative: 0.01% Sodium Azide
Constituents: 40% Glycerol, 0.05% BSA, 0.15M Sodium chloride, 50mM Tris glycine, pH 7.4
Purity
Tissue culture supernatant
Clonality
Monoclonal
Clone number
EP568Y
Isotype
IgG
Research areas
Cancer >> Cancer Metabolism >> Response to hypoxia
Cancer >> Signal transduction >> Nuclear signaling >> SMAD family
Stem Cells >> Signaling Pathways >> TGF beta >> Cytoplasmic
Epigenetics and Nuclear Signaling >> Nuclear Signaling Pathways >> SMADs
Signal Transduction >> Signaling Pathway >> Nuclear Signaling >> SMADs
Applications
Show applications key
Our Abpromise guarantee covers the use of ab40854 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
WB: 1/5000Detects a band of approximately 55 kDa (predicted molecular weight: 48 kDa).
ICC
ICC: 1/100 - 1/250.ICC: 1/100 - 1/250.
IHC-P: 1/100 - 1/250.
IP
IP: 1/50IP: 1/50
sELISA: Use a concentration of 0.5 µg/ml Can be paired for Sandwich ELISA with Mouse monoclonal [AF9F7] to Smad3 (ab75512). (For sandwich ELISA, use this antibody as Detection at 0.5 µg/ml with Mouse monoclonal [AF9F7] to Smad3 (ab75512) as Capture.)
Target
Function
Receptor-regulated SMAD (R-SMAD) that is an intracellular signal transducer and transcriptional modulator activated by TGF-beta (transforming growth factor) and activin type 1 receptor kinases. Binds the TRE element in the promoter region of many genes that are regulated by TGF-beta and, on formation of the SMAD3/SMAD4 complex, activates transcription. Also can form a SMAD3/SMAD4/JUN/FOS complex at the AP-1/SMAD site to regulate TGF-beta-mediated transcription. Has an inhibitory effect on wound healing probably by modulating both growth and migration of primary keratinocytes and by altering the TGF-mediated chemotaxis of monocytes. This effect on wound healing appears to be hormone-sensitive. Regulator of chondrogenesis and osteogenesis and inhibits early healing of bone fractures.
Involvement in disease
Defects in SMAD3 may be a cause of colorectal cancer (CRC) [MIM:114500].
Sequence similarities
Belongs to the dwarfin/SMAD family.
Contains 1 MH1 (MAD homology 1) domain.
Contains 1 MH2 (MAD homology 2) domain.
Domain
The MH1 domain is required for DNA binding. Also binds zinc ions which are necessary for the DNA binding.
The MH2 domain is required for both homomeric and heteromeric interactions and for transcriptional regulation. Sufficient for nuclear import.
The linker region is required for the TGFbeta-mediated transcriptional activity and acts synergistically with the MH2 domain.
Post-translational
modifications
Phosphorylated on serine and threonine residues. Enhanced phosphorylation in the linker region on Thr-179, Ser-204 and Ser-208 on EGF AND TGF-beta treatment. Ser-208 is the main site of MAPK-mediated phosphorylation. CDK-mediated phosphorylation occurs in a cell-cycle dependent manner and inhibits both the transcriptional activity and antiproliferative functions of SMAD3. This phosphorylation is inhibited by flavopiridol. Maximum phosphorylation at the G(1)/S junction. Also phosphorylated on serine residues in the C-terminal SXS motif by TGFBR1 and ACVR1. TGFBR1-mediated phosphorylation at these C-terminal sites is required for interaction with SMAD4, nuclear location and transactivational activity, and appears to be a prerequisite for the TGF-beta mediated phosphorylation in the linker region. Dephosphorylated in the C-terminal SXS motif by PPM1A. This dephosphorylation disrupts the interaction with SMAD4, promotes nuclear export and terminates TGF-beta-mediated signaling.
Acetylation in the nucleus by EP300 in the MH2 domain regulates positively its transcriptional activity and is enhanced by TGF-beta.
Cellular localization
Cytoplasm. Nucleus. Cytoplasmic and nuclear in the absence of TGF-beta. On TGF-beta stimulation, migrates to the nucleus when complexed with SMAD4. Through the action of the phosphatase PPM1A, released from the SMAD2/SMAD4 complex, and exported out of the nucleus by interaction with RANBP1. Co-localizes with LEMD3 at the nucleus inner membrane. MAPK-mediated phosphorylation appears to have no effect on nuclear import.
Target information above from: UniProt accessionP84022
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Alternative names
- DKFZP586N0721 antibody
- DKFZp686J10186 antibody
- hMAD 3 antibody
see all
Database links
- Entrez Gene: 4088 Human
- Entrez Gene: 17127 Mouse
- Entrez Gene: 25631 Rat
- Omim: 603109 Human
- SwissProt: P84022 Human
- SwissProt: Q8BUN5 Mouse
- SwissProt: P84025 Rat
- Unigene: 36915 Human
see all
Anti-Smad3 antibody [EP568Y] images:
Sandwich ELISA - Smad3 antibody [EP568Y] (ab40854)
![Sandwich ELISA - Smad3 antibody [EP568Y] (ab40854)](/ps/datasheet/images/40/ab40854/Smad3-Primary-antibodies-ab40854-1.jpg)
Standard Curve for Smad3 (Analyte: Smad3 protein (His tag) (ab89353)); dilution range 1pg/ml to 1µg/ml using Capture Antibody Mouse monoclonal [AF9F7] to Smad3 (ab75512) at 5µg/ml and Detector Antibody Rabbit monoclonal [EP568Y] to Smad3 (ab40854) at 0.5µg/ml.
Western blot - Smad3 antibody [EP568Y] (ab40854)
![Western blot - Smad3 antibody [EP568Y] (ab40854)](/ps/datasheet/Images/40/ab40854/ab40854_1.jpg)
Anti-Smad3 antibody [EP568Y] (ab40854) at 1/5000 dilution + Jurkat cell lysate at 10 µg
Predicted band size : 48 kDa
Observed band size : 55 kDa (why is the actual band size different from the predicted?)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Smad3 antibody [EP568Y] (ab40854)
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Smad3 antibody [EP568Y] (ab40854)](/ps/datasheet/Images/40/ab40854/ab40854_2.jpg)
ab40854 at a 1:100 dilution staining Smad3 in human prostate carcinoma tissue.
Protocols
References for Anti-Smad3 antibody [EP568Y] (ab40854)
This product has been referenced in:
- González AMet al. Assessment of the protective effects of oral tocotrienols in arginine chronic-like pancreatitis. Am J Physiol Gastrointest Liver Physiol 301:G846-55 (2011). WB; Rat.Read more (PubMed: 21852363) »
See 1 publication for this product
Publishing research using ab40854? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"