Rabbit Smad3 Polyclonal Antibody | anti-Smad3 antibody

Smad3 Antibody

Cat. Num. AAA9613520

• Gene Names: SMAD3; LDS3; LDS1C; MADH3; JV15-2; HSPC193; HsT17436
• Reactivity: Human, Mouse, Rat; Predicted Reactivity: Pig (100%), Bovine (100%), Horse (100%), Sheep (100%), Rabbit (100%), Dog (100%), Chicken (100%), Xenopus (86%)
• Applications: Immunofluorescence, Immunocytochemistry, ELISA
• Purity: The antiserum was purified by peptide affinity chromatography using SulfoLink Coupling Resin
• Synonyms: Smad3; Polyclonal Antibody; Smad3 Antibody; DKFZP586N0721; DKFZp686J10186; hMAD 3; hMAD-3; hSMAD3; HSPC193; HST17436; JV15 2; JV15-2; JV152; LDS1C; LDS3; MAD (mothers against decapentaplegic Drosophila) homolog 3; MAD homolog 3; Mad homolog JV15 2; Mad protein homolog; MAD; mothers against decapentaplegic homolog 3; Mad3; MADH 3; MADH3; MGC60396; Mothers against decapentaplegic homolog 3; Mothers against DPP homolog 3; SMA and MAD related protein 3; SMAD 3; SMAD; SMAD family member 3; mothers against DPP homolog 3; SMAD3_HUMAN; anti-Smad3 antibody

• Host: Rabbit
• Reactivity: Human, Mouse, Rat; Predicted Reactivity: Pig (100%), Bovine (100%), Horse (100%), Sheep (100%), Rabbit (100%), Dog (100%), Chicken (100%), Xenopus (86%)
• Clonality: Polyclonal
• Isotype: Rabbit IgG
• Specificity: Smad3 Antibody detects endogenous levels of total Smad3.
• Purity/Purification: The antiserum was purified by peptide affinity chromatography using SulfoLink Coupling Resin
• Form/Format: Liquid. Rabbit IgG in phosphate buffered saline, pH7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.
• Concentration: 1mg/ml (varies by lot)

• Applicable Applications for anti-Smad3 antibody: Immunofluorescence (IF), Immunocytochemistry (ICC), Peptide ELISA (EIA)
• Application Notes: IF/ICC: 1:100-1:500; Peptide ELISA: 1:20,000-1:40,000
• Immunogen: A synthesized peptide derived from human Smad3.
• Conjugation: Unconjugated
• Fragment: Fab fragment
• 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. Phosphorylation at Ser-418 by CSNK1G2/CK1 promotes ligand-dependent ubiquitination and subsequent proteasome degradation, thus inhibiting SMAD3-mediated TGF-beta responses. Phosphorylated by PDPK1. Acetylation in the nucleus by EP300 in the MH2 domain regulates positively its transcriptional activity and is enhanced by TGF-beta.Poly-ADP-ribosylated by PARP1 and PARP2. ADP-ribosylation negatively regulates SMAD3 transcriptional responses during the course of TGF-beta signaling.Ubiquitinated. Monoubiquitinated, leading to prevent DNA-binding. Deubiquitination by USP15 alleviates inhibition and promotes activation of TGF-beta target genes. Ubiquitinated by RNF111, leading to its degradation: only SMAD3 proteins that are 'in use' are targeted by RNF111, RNF111 playing a key role in activating SMAD3 and regulating its turnover (By similarity). Undergoes STUB1-mediated ubiquitination and degradation.
• Subunit Structure: Monomer; in the absence of TGF-beta. Homooligomer; in the presence of TGF-beta. Heterotrimer; forms a heterotrimer in the presence of TGF-beta consisting of two molecules of C-terminally phosphorylated SMAD2 or SMAD3 and one of SMAD4 to form the transcriptionally active SMAD2/SMAD3-SMAD4 complex. Interacts with TGFBR1. Part of a complex consisting of AIP1, ACVR2A, ACVR1B and SMAD3. Interacts with AIP1, TGFB1I1, TTRAP, FOXL2, PML, PRDM16, HGS, WWP1 and SNW1. Interacts (via MH2 domain) with CITED2 (via C-terminus). Interacts with NEDD4L; the interaction requires TGF-beta stimulation. Interacts (via MH2 domain) with ZFYVE9. Interacts with HDAC1, TGIF and TGIF2, RUNX3, CREBBP, SKOR1, SKOR2, SNON, ATF2, SMURF2 and TGFB1I1. Interacts with DACH1; the interaction inhibits the TGF-beta signaling. Forms a complex with SMAD2 and TRIM33 upon addition of TGF-beta. Found in a complex with SMAD3, RAN and XPO4. Interacts in the complex directly with XPO4. Interacts (via MH2 domain) with LEMD3; the interaction represses SMAD3 transcriptional activity through preventing the formation of the heteromeric complex with SMAD4 and translocation to the nucleus. Interacts with RBPMS. Interacts (via MH2 domain) with MECOM. Interacts with WWTR1 (via its coiled-coil domain). Interacts (via the linker region) with EP300 (C-terminal); the interaction promotes SMAD3 acetylation and is enhanced by TGF-beta phosphorylation in the C-terminal of SMAD3. This interaction can be blocked by competitive binding of adenovirus oncoprotein E1A to the same C-terminal site on EP300, which then results in partially inhibited SMAD3/SMAD4 transcriptional activity. Interacts with SKI; the interaction represses SMAD3 transcriptional activity. Component of the multimeric complex SMAD3/SMAD4/JUN/FOS which forms at the AP1 promoter site; required for synergistic transcriptional activity in response to TGF-beta. Interacts (via an N-terminal domain) with JUN (via its basic DNA binding and leucine zipper domains); this interaction is essential for DNA binding and cooperative transcriptional activity in response to TGF-beta. Interacts with PPM1A; the interaction dephosphorylates SMAD3 in the C-terminal SXS motif leading to disruption of the SMAD2/3-SMAD4 complex, nuclear export and termination of TGF-beta signaling. Interacts (dephosphorylated form via the MH1 and MH2 domains) with RANBP3 (via its C-terminal R domain); the interaction results in the export of dephosphorylated SMAD3 out of the nucleus and termination of the TGF-beta signaling. Interacts with MEN1. Interacts with IL1F7. Interaction with CSNK1G2. Interacts with PDPK1 (via pHdomain). Interacts with DAB2; the interactions are enhanced upon TGF-beta stimulation. Interacts with USP15. Interacts with PPP5C; the interaction decreases SMAD3 phosphorylation and protein levels. Interacts with LDLRAD4 (via the SMAD interaction motif). Interacts with PMEPA1. Interacts with ZC3H3 (By similarity). Interacts with ZNF451. Identified in a complex that contains at least ZNF451, SMAD2, SMAD3 and SMAD4. Interacts with ZFHX3. Interacts weakly with ZNF8. Interacts (when phosphorylated) with RNF111; RNF111 acts as an enhancer of the transcriptional responses by mediating ubiquitination and degradation of SMAD3 inhibitors (By similarity). Interacts with STUB1, HSPA1A, HSPA1B, HSP90AA1 and HSP90AB1. Interacts (via MH2 domain) with ZMIZ1 (via SP-RING-type domain); in the TGF-beta signaling pathway increases the activity of the SMAD3/SMAD4 transcriptional complex.
• Similarity: 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.Belongs to the dwarfin/SMAD family.
• Subcellular Location: Cytoplasm. Nucleus.; Note: Cytoplasmic and nuclear in the absence of TGF-beta. On TGF-beta stimulation, migrates to the nucleus when complexed with SMAD4 (PubMed:15799969). Through the action of the phosphatase PPM1A, released from the SMAD2/SMAD4 complex, and exported out of the nucleus by interaction with RANBP1 (PubMed:16751101, PubMed:19289081). Co-localizes with LEMD3 at the nucleus inner membrane (PubMed:15601644). MAPK-mediated phosphorylation appears to have no effect on nuclear import (PubMed:19218245). PDPK1 prevents its nuclear translocation in response to TGF-beta (PubMed:17327236).
• Preparation and Storage: Store at -20 degree C. Stable for 12 months from date of receipt.

Immunofluorescence (IF)/Immunocytochemistry (ICC)

(Staining Hela cells by IF/ICC. The samples were fixed with PFA and permeabilized in 0.1% Triton X-100,then blocked in 10% serum for 45 minutes at 25 degree C. Samples were then incubated with primary Ab(AF6365 1:200) and mouse anti-beta tubulin Ab(T0023 1:200) for 1 hour at 37 degree C. An AlexaFluor594 conjugated goat anti-rabbit IgG(H+L) Ab(Red) and an AlexaFluor488 conjugated goat anti-mouse IgG(H+L) Ab(Green) were used as the secondary antibody.The nuclear counter stain is DAPI(blue).)

Related Product Information for anti-Smad3 antibody

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. Positively regulates PDPK1 kinase activity by stimulating its dissociation from the 14-3-3 protein YWHAQ which acts as a negative regulator.

NCBI and Uniprot Product Information

• NCBI GI #: 223029440
• NCBI GeneID: 4088
• NCBI Accession #: NP_001138574.1
• NCBI GenBank Nucleotide #: NM_001145102.1
• UniProt Accession #: P84022; O09064; O09144; O14510; O35273; Q92940; Q93002; A8K4B6; B7Z4Z5; B7Z6M9; B7Z9Q2; F5H383
• Molecular Weight: 48,081 Da
• NCBI Official Full Name: mothers against decapentaplegic homolog 3 isoform 2
• NCBI Official Synonym Full Names: SMAD family member 3
• NCBI Official Symbol: SMAD3
• NCBI Official Synonym Symbols: LDS3; LDS1C; MADH3; JV15-2; HSPC193; HsT17436
• NCBI Protein Information: mothers against decapentaplegic homolog 3; mad3; hMAD-3; hSMAD3; MAD homolog 3; mad homolog JV15-2; mad protein homolog; mothers against DPP homolog 3; SMA- and MAD-related protein 3; SMAD, mothers against DPP homolog 3; MAD, mothers against decapentapleg
• UniProt Protein Name: Mothers against decapentaplegic homolog 3
• Protein Family: Mothers against decapentaplegic
• UniProt Gene Name: SMAD3
• UniProt Synonym Gene Names: MADH3; MAD homolog 3; Mad3; Mothers against DPP homolog 3; hMAD-3; SMAD 3; Smad3; hSMAD3
• UniProt Entry Name: SMAD3_HUMAN

NCBI Description

The protein encoded by this gene belongs to the SMAD, a family of proteins similar to the gene products of the Drosophila gene 'mothers against decapentaplegic' (Mad) and the C. elegans gene Sma. SMAD proteins are signal transducers and transcriptional modulators that mediate multiple signaling pathways. This protein functions as a transcriptional modulator activated by transforming growth factor-beta and is thought to play a role in the regulation of carcinogenesis. [provided by RefSeq, Apr 2009]

Uniprot Description

SMAD3: transcription factor phosphorylated and activated by TGF-beta-type receptors. A receptor-regulated Smad (R-smad). Binds directly to consensus DNA-binding elements in the promoters of target genes. In mouse required for establishemnt of the mucosal immune response and proper development of skeleton.

Protein type: DNA-binding; Transcription factor; Nuclear receptor co-regulator

Chromosomal Location of Human Ortholog: 15q22.33

Cellular Component: nucleoplasm; transcription factor complex; nuclear chromatin; cytoplasm; plasma membrane; nuclear inner membrane; cytosol; nucleus; receptor complex

Molecular Function: identical protein binding; ubiquitin binding; protein homodimerization activity; zinc ion binding; chromatin DNA binding; beta-catenin binding; protein kinase binding; transcription factor binding; phosphatase binding; transforming growth factor beta receptor, pathway-specific cytoplasmic mediator activity; collagen binding; protein binding; sequence-specific DNA binding; ubiquitin protein ligase binding; double-stranded DNA binding; bHLH transcription factor binding; transforming growth factor beta receptor binding; transcription factor activity

Biological Process: developmental growth; positive regulation of positive chemotaxis; positive regulation of transcription, DNA-dependent; activin receptor signaling pathway; paraxial mesoderm morphogenesis; embryonic pattern specification; regulation of transforming growth factor beta receptor signaling pathway; transforming growth factor beta receptor signaling pathway; positive regulation of stress fiber formation; embryonic foregut morphogenesis; negative regulation of mitotic cell cycle; negative regulation of osteoblast differentiation; cell cycle arrest; regulation of striated muscle development; pericardium development; somitogenesis; transcription, DNA-dependent; embryonic cranial skeleton morphogenesis; osteoblast development; regulation of transcription from RNA polymerase II promoter; positive regulation of chondrocyte differentiation; mesoderm formation; negative regulation of osteoblast proliferation; positive regulation of transcription from RNA polymerase II promoter; negative regulation of protein catabolic process; endoderm development; negative regulation of apoptosis; evasion of host defenses by virus; wound healing; T cell activation; negative regulation of transcription from RNA polymerase II promoter; primary microRNA processing; regulation of epithelial cell proliferation; positive regulation of focal adhesion formation; negative regulation of protein amino acid phosphorylation; ureteric bud development; transport; positive regulation of transforming growth factor-beta3 production; thyroid gland development; positive regulation of transcription factor import into nucleus; heart looping; caspase activation; transcription initiation from RNA polymerase II promoter; intercellular junction assembly and maintenance; regulation of transforming growth factor-beta2 production; regulation of immune response; regulation of binding; protein stabilization; in utero embryonic development; positive regulation of bone mineralization; liver development; SMAD protein complex assembly; immune system development; positive regulation of interleukin-1 beta production; negative regulation of inflammatory response; response to hypoxia; gene expression; immune response; negative regulation of cell growth; negative regulation of transforming growth factor beta receptor signaling pathway; positive regulation of cell migration

Disease: Loeys-dietz Syndrome 3