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Products:Signal Transduction >> Signaling Pathway >> Nuclear Signaling >> SMADs
Overview
Product name
Anti-Smad3 (phospho S204) antibody
See all Smad3 products (21) ...
Description
Rabbit polyclonal to Smad3 (phospho S204)
Specificity
This antibody detects endogenous levels of Smad3 only when phosphorylated at serine 204.
Tested applications
WB, ELISAmore details
Cross reactivity
Reacts with
Human
Immunogen
A synthesized phosphopeptide derived from human Smad3 around the phosphorylation site of serine 204 (AGSPPN).
Positive control
Extracts from NIH/3T3 cells
Properties
Form
Liquid
Storage instructions
Store at -20°C. Stable for 12 months at -20°C
Storage buffer
Preservative: 0.02% Sodium Azide
Constituents: 50% Glycerol, PBS (without Mg+2 and Ca+2), 150mM Sodium chloride, pH 7.4
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Purification notes
The antibody was affinity purified from rabbit antiserum by affinity chromatography using epitope specific phosphopeptide. The antibody against non phosphopeptide was removed by chromatography using non phosphopeptide corresponding to the phosphorylation site.
Clonality
Polyclonal
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
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Western blot - Smad3 (phospho S204) antibody (ab63402)
(enlarge)
Applications
Show applications key
Our Abpromise guarantee covers the use of ab63402 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
WB: 1/500 - 1/1000.Detects a band of approximately 48 kDa (predicted molecular weight: 48 kDa).
ELISA
ELISA: 1/20000ELISA: 1/20000
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
- Omim: 603109 Human
- SwissProt: P84022 Human
- Unigene: 36915 Human
Anti-Smad3 (phospho S204) antibody images:
Western blot - Smad3 (phospho S204) antibody (ab63402)
All lanes : Anti-Smad3 (phospho S204) antibody (ab63402) at 1/500 dilution
Lane 1 : Extracts from NIH/3T3 cells,
treated with Serum (20%, 15mins)
Lane 2 : Extracts from NIH/3T3 cells,
treated with Serum (20%, 15mins) plus phospho peptide
Predicted band size : 48 kDa
Observed band size : 48 kDa
The amount of positive control loading for the WB is 5-30 ug of total protein. The amount of the peptide is 5-10 ug.
References for Anti-Smad3 (phospho S204) antibody (ab63402)
ab63402 has not yet been referenced specifically in any publications.
Publishing research using ab63402? 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"