• Western blot analysis of extracts of C6 cells, using Phospho-Smad3-S423/S425 rabbit monoclonal antibody (STJ11102566) at 1:1000 dilution. C6 cells were treated by PMA/TPA (200 nM) at 37 °C for 10 minutes. Secondary antibody: HRP Goat Anti-rabbit IgG (H+L) (STJS000856) at 1:10000 dilution. Lysates/proteins: 25 Mu g per lane. Blocking buffer: 3% non-fat dry milk in TBST. Detection: ECL Basic Kit. Exposure time: 60s.
  • Immunohistochemistry analysis of paraffin-embedded mouse kidney using Phospho-Smad3-S423/S425 rabbit monoclonal antibody (STJ11102566) at dilution of 1:200 (40x lens). Perform high pressure antigen retrieval with 10 mM citrate buffer pH 6. 0 before commencing with immunohistochemistry staining protocol.
  • Immunohistochemistry analysis of paraffin-embedded rat ovary using Phospho-Smad3-S423/S425 rabbit monoclonal antibody (STJ11102566) at dilution of 1:200 (40x lens). Perform high pressure antigen retrieval with 10 mM citrate buffer pH 6. 0 before commencing with immunohistochemistry staining protocol.
  • Immunohistochemistry analysis of paraffin-embedded human colon carcinoma using Phospho-Smad3-S423/S425 rabbit monoclonal antibody (STJ11102566) at dilution of 1:200 (40x lens). Perform high pressure antigen retrieval with 10 mM citrate buffer pH 6. 0 before commencing with immunohistochemistry staining protocol.
  • Immunohistochemistry analysis of paraffin-embedded rat lung using Phospho-Smad3-S423/S425 rabbit monoclonal antibody (STJ11102566) at dilution of 1:200 (40x lens). Perform high pressure antigen retrieval with 10 mM citrate buffer pH 6. 0 before commencing with immunohistochemistry staining protocol.
  • Immunohistochemistry analysis of paraffin-embedded human breast cancer using Phospho-Smad3-S423/S425 rabbit monoclonal antibody (STJ11102566) at dilution of 1:200 (40x lens). Perform high pressure antigen retrieval with 10 mM citrate buffer pH 6. 0 before commencing with immunohistochemistry staining protocol.

Anti-Phospho-Smad3-S423/S425 antibody [S6MR] (STJ11102566)

SKU:
STJ11102566

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Host: Rabbit
Applications: WB/IHC
Reactivity: Human/Mouse/Rat
Note: STRICTLY FOR FURTHER SCIENTIFIC RESEARCH USE ONLY (RUO). MUST NOT TO BE USED IN DIAGNOSTIC OR THERAPEUTIC APPLICATIONS.
Short Description: Rabbit monoclonal antibody anti-Phospho-Smad3-S423/S425 is suitable for use in Western Blot and Immunohistochemistry research applications.
Clonality: Monoclonal
Clone ID: S6MR
Conjugation: Unconjugated
Isotype: IgG
Formulation: PBS with 0.02% Sodium Azide, 0.05% BSA, 50% Glycerol, pH7.3.
Purification: Affinity purification
Dilution Range: WB 1:500-1:2000
IHC-P 1:50-1:200
Storage Instruction: Store at-20°C for up to 1 year from the date of receipt, and avoid repeat freeze-thaw cycles.
Gene Symbol: SMAD3
Gene ID: 4088
Uniprot ID: SMAD3_HUMAN
Immunogen: A synthetic phosphorylated peptide around S423 & S425 of human Smad3 (P84022).
Immunogen Sequence: SSVS
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. Undergoes STUB1-mediated ubiquitination and degradation.
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. Positively regulates PDPK1 kinase activity by stimulating its dissociation from the 14-3-3 protein YWHAQ which acts as a negative regulator.
Protein Name Mothers Against Decapentaplegic Homolog 3
Mad Homolog 3
Mad3
Mothers Against Dpp Homolog 3
Hmad-3
Jv15-2
Smad Family Member 3
Smad 3
Smad3
Hsmad3
Database Links Reactome: R-HSA-1181150
Reactome: R-HSA-1502540
Reactome: R-HSA-2173788
Reactome: R-HSA-2173789
Reactome: R-HSA-2173795
Reactome: R-HSA-2173796
Reactome: R-HSA-3304356
Reactome: R-HSA-3311021
Reactome: R-HSA-3315487
Reactome: R-HSA-3656532
Reactome: R-HSA-5689880
Reactome: R-HSA-8941855
Reactome: R-HSA-8952158
Reactome: R-HSA-9008059
Reactome: R-HSA-9013695
Reactome: R-HSA-9615017
Reactome: R-HSA-9617828
Reactome: R-HSA-9735871
Reactome: R-HSA-9754189
Reactome: R-HSA-9796292
Reactome: R-HSA-9823730
Cellular Localisation 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
Pdpk1 Prevents Its Nuclear Translocation In Response To Tgf-Beta
Localized Mainly To The Nucleus In The Early Stages Of Embryo Development With Expression Becoming Evident In The Cytoplasm Of The Inner Cell Mass At The Blastocyst Stage
Alternative Antibody Names Anti-Mothers Against Decapentaplegic Homolog 3 antibody
Anti-Mad Homolog 3 antibody
Anti-Mad3 antibody
Anti-Mothers Against Dpp Homolog 3 antibody
Anti-Hmad-3 antibody
Anti-Jv15-2 antibody
Anti-Smad Family Member 3 antibody
Anti-Smad 3 antibody
Anti-Smad3 antibody
Anti-Hsmad3 antibody
Anti-SMAD3 antibody
Anti-MADH3 antibody

Information sourced from Uniprot.org

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