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Products:Signal Transduction >> Signaling Pathway >> Nuclear Signaling >> SMADs
Overview
Product name
Anti-Smad3 (phospho S423 + S425) antibody
See all Smad3 products (21) ...
Description
Rabbit polyclonal to Smad3 (phospho S423 + S425)
Specificity
ab51451 is specific for the phosphorylated form of the protein although minimal reactivity with nonphosphorylated human Smad3 can be observed by ELISA and WB. ab51451 also reacts with phosphorylated Smad1 and Smad5 but negligible reactivity is seen against other phosphorylated Smad family members.
Tested applications
IHC-P, WB, ELISA, ICC/IFmore details
Cross reactivity
Reacts with
Mouse, Chicken, Human
Predicted to work with
Rat, Cow, Pig, Xenopus laevis, Zebrafish
Immunogen
Synthetic peptide: PSIRCSSpVSp, corresponding to amino acids 417-425 of Human Smad3
PSIRCSSVS
Properties
Form
Liquid
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage buffer
Preservative: 0.01% Sodium Azide
Constituents: 0.15M Sodium chloride, 0.02M Potassium phosphate, pH 7.2
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Purification notes
ab51451 was affinity purified from monospecific antiserum by immunoaffinity purification. Antiserum was first purified against the phosphorylated form of the immunizing peptide. The resultant affinity purified antibody was then cross adsorbed against the non-phosphorylated form of the immunizing peptide.
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
Applications
Show applications key
Our Abpromise guarantee covers the use of ab51451 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
IHC-P: Use at an assay dependent dilution. (PubMed: 20519446)
WB: 1/2000 - 1/20000.Predicted molecular weight: 48 kDa.(Stimulation with 2 ng/ml TGF-beta for 1 hour is suggested.)
ELISA: 1/25000 - 1/100000.
ICC/IF: Use at an assay dependent dilution.
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: 397260 Pig
- Entrez Gene: 25631 Rat
- Omim: 603109 Human
- SwissProt: P84023 Chicken
- SwissProt: P84022 Human
- SwissProt: Q8BUN5 Mouse
see all
Anti-Smad3 (phospho S423 + S425) antibody images:
Western blot - Smad3 (phospho S423 + S425) antibody (ab51451)
All lanes : Anti-Smad3 (phospho S423 + S425) antibody (ab51451) at 1/2000 dilution
Lane 1 : MEF lysate (control)
Lane 2 : MEF lysate, after stimulation with 2 ng/ml of TGF-ß for 1 hour
Lane 3 : MEF-/-Smad3 lysate (control)
Lane 4 : MEF-/-Smad3 lysate, after stimulation with 2 ng/ml of TGF-ß for one hour
Lane 5 : Hep3B lysate (control)
Lane 6 : Hep3B lysate, after stimulation with 2 ng/ml of TGF-ß for one hour
Lane 7 : HEK 293 lysate (control)
Predicted band size : 48 kDa
Image courtesy of Ying Zhang, NIH, CCR, Bethesda, MD.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Smad3 (phospho S423 + S425) antibody (ab51451)
ab51451 at 2.5 µg/ml staining epidermal keratinocytes at 40X. This image shows strong nuclear staining with localization of the antibody as the precipated red siganl and a hematoxylin purple nuclear counterstain.
References for Anti-Smad3 (phospho S423 + S425) antibody (ab51451)
This product has been referenced in:
- Avraham Tet al. Radiation therapy causes loss of dermal lymphatic vessels and interferes with lymphatic function by TGF-beta1-mediated tissue fibrosis. Am J Physiol Cell Physiol 299:C589-605 (2010). IHC-P; Mouse.Read more (PubMed: 20519446) »
- Avraham Tet al. Blockade of Transforming Growth Factor-{beta}1 Accelerates Lymphatic Regeneration during Wound Repair. Am J Pathol 177:3202-14 (2010). IHC-P; Mouse.Read more (PubMed: 21056998) »
See all 5 publications for this product
Publishing research using ab51451? 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"