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Anti-Phospho-Smad3 (S423 + S425) Rabbit Monoclonal Antibody

All lanes use the Antibody at 1:1K dilution for 1 hour at room temperature.
All lanes use the Antibody at 1:1K dilution for 1 hour at room temperature.
All lanes use the Antibody at 1:1K dilution for 1 hour at room temperature.
All lanes use the Antibody at 1:1K dilution for 1 hour at room temperature.
Western blot analysis of Phospho-Smad3 (S423/S425) expression in A549 cell lysate treated with TGF-?1.
Immunohistochemical analysis of paraffin-embedded human gastric adenocarcinoma, using Phospho-Smad3 (S423 + S425) Antibody.
Immunofluorescent analysis of A549 cells treated with TGFβ , using Phospho-Smad3 (S423 + S425) Antibody.

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Product Information
Application Applications Legend: E=ELISA WB=Western Blotting IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin) IP=Immunoprecipitation IF=Immunofluorescence IC=Immunochemistry ICC=Immunocytochemistry FC=Flow Cytometry DB=Dot Blot	WB, IHC, IF, ICC
Primary Accession	P84022
Host	Rabbit
Isotype	Rabbit IgG
Reactivity	Human, Mouse
Clonality	Monoclonal
Format	Liquid
Description	Anti-Phospho-Smad3 (S423 + S425) Rabbit Monoclonal Antibody . Tested in WB, IHC, ICC/IF applications. This antibody reacts with Human, Mouse.

Additional Information
Gene ID	4088
Other Names	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, SMAD3, MADH3
Calculated MW	48081 Da
Application Details	WB 1:500-1:2000 IHC 1:50-1:200 ICC/IF 1:50-1:200
Subcellular 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. PDPK1 prevents its nuclear translocation in response to TGF-beta.
Source	Eukaryota
Contents	Rabbit IgG in phosphate buffered saline, pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol, 0.4-0.5mg/ml BSA.
Clone Names	Clone: AHO-19
Immunogen	A synthesized peptide derived from human Phospho-Smad3 (S423 + S425)
Purification	Affinity-chromatography
Storage	Store at -20°C for one year. For short term storage and frequent use, store at 4°C for up to one month. Avoid repeated freeze-thaw cycles.

For Research Use Only. Not For Use In Diagnostic Procedures.

Protein Information
Name	SMAD3 (HGNC:6769)
Synonyms	MADH3
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.
Cellular 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, PubMed:21145499). 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). 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 (By similarity). {ECO:0000250\|UniProtKB:Q8BUN5, ECO:0000269\|PubMed:15601644, ECO:0000269\|PubMed:15799969, ECO:0000269\|PubMed:16751101, ECO:0000269\|PubMed:17327236, ECO:0000269\|PubMed:19218245, ECO:0000269\|PubMed:19289081, ECO:0000269\|PubMed:21145499}

Research Areas

Application Protocols

Provided below are standard protocols that you may find useful for product applications.

Western Blot Protocol

Blocking Peptides Protocol

Dot Blot Protocol

Immunohistochemistry Protocol

Immunofluorescence Protocol

Immunoprecipitation Protocol

Flow Cytomety Protocol

Cell Culture Protocol