Recombinant Human SMAD3 Protein (N-6His-Flag)

Name: Recombinant Human SMAD3 Protein (N-6His-Flag)
Brand: Beta LifeScience
SKU: BL-1854NP
Price: 233.0 USD
Availability: InStock

BL-1854NP: Greater than 85% as determined by reducing SDS-PAGE. (QC verified)

Collections: Other recombinant proteins, Recombinant proteins

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Product Overview

Description	Recombinant Human Mothers Against Decapentaplegic Homolog 3 is produced by our E.coli expression system and the target gene encoding Ser2-Ser425 is expressed with a 6His, Flag tag at the N-terminus.
Accession	P84022
Synonym	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
Gene Background	Mothers against decapentaplegic homolog 3(SMAD3) is a cytoplasm protein which belongs to the dwarfin/SMAD family. Smad proteins undergo rapid nuclear translocation upon stimulation by transforming growth factor and in so doing transduce the signal into the nucleus. Receptor-regulated SMAD is an intracellular signal transducer and transcriptional modulator activated by TGF-beta and activin type 1 receptor kinases. SMAD3 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. It also can form a SMAD3/SMAD4/JUN/FOS complex at the AP-1/SMAD site to regulate TGF-beta-mediated transcription. SMAD3 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.
Molecular Mass	50.5 KDa
Apmol Mass	50-60 KDa, reducing conditions
Formulation	Supplied as a 0.2 μm filtered solution of 20mM Tris-HCl, 500mM NaCl, 10% Glycerol, 2mM EDTA, pH 8.0.
Endotoxin	Less than 0.1 ng/µg (1 EU/µg) as determined by LAL test.
Purity	Greater than 85% as determined by reducing SDS-PAGE. (QC verified)
Biological Activity	Not tested
Reconstitution
Storage	Lyophilized protein should be stored at ≤ -20°C, stable for one year after receipt. Reconstituted protein solution can be stored at 2-8°C for 2-7 days. Aliquots of reconstituted samples are stable at ≤ -20°C for 3 months.
Shipping	The product is shipped on dry ice/polar packs. Upon receipt, store it immediately at the temperature listed below.
Usage	For Research Use Only

Target Details

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. Positively regulates PDPK1 kinase activity by stimulating its dissociation from the 14-3-3 protein YWHAQ which acts as a negative regulator.
Subcellular Location	Cytoplasm. Nucleus.
Protein Families	Dwarfin/SMAD family
Database References	HGNC: 6769 OMIM: 114500 KEGG: hsa:4088 STRING: 9606.ENSP00000332973 UniGene: Hs.727986
Associated Diseases	Colorectal cancer (CRC); Loeys-Dietz syndrome 3 (LDS3)

Gene Functions References

Study established a relationship between OCT4 and SMAD3 heterodimers formation and Snail, Slug, and CXCL13 transcription promotion mediating breast cancer progression. PMID: 29526821
Study results using gene editing indicate the cancer-promoting role of Smad3 T179 phosphorylation in the human triple-negative breast cancer cells. PMID: 30251686
Downregulation of miR-637 promotes proliferation and migration of fibroblasts by targeting Smad3 in keloids. PMID: 29845237
The findings of the present study indicated that miR326 inhibited endometrial fibrosis by suppressing the TGFbeta1/Smad3 signaling pathway, suggesting that miR326 may be a prognostic biomarker and therapeutic target for Intrauterine adhesion (IUA). PMID: 29956752
Study validated a specific model prediction that SMAD3 regulates Huntington's disease (HD)-related gene expression changes. Also, results found CAG repeat length-dependent changes in the genomic occupancy of SMAD3 and confirmed the model's prediction that many SMAD3 target genes are downregulated early in HD. PMID: 29581148
The SMAD3 rs12901499 polymorphism may be involved in the development of knee osteoarthritis. Larger studies with more diverse ethnic populations are needed to confirm these result. PMID: 29315792
NLRC5 may act as a key mediator in renal fibroblast activation and fibrogenesis PMID: 29608899
The SMAD3 SNP rs12901499 GA genotype and G variant may increase the risk of hip osteoarthritis in Chinese Han patients. PMID: 29310478
Positive cooperativity of Smad3 and STAT3 during epithelial-mesenchymal transition [Review]. PMID: 29140406
CXCL12 activates the MEKK1/JNK signaling pathway, which in turn initiates SMAD3 phosphorylation, its translocation to nuclei, and recruitment of SMAD3 to the CTGF promoter, which ultimately induces CTGF expression in human lung fibroblasts. PMID: 29499695
these results indicated that Bone marrow-derived mesenchymal stem cells -conditioned medium suppressed the epithelial-mesenchymal transition which might be associated with TGF-B1/Smad3. This study provides the theoretical basis for the research of the mechanisms responsible for pulmonary disease. PMID: 29207055
The present findings indicate that RACK1 silencing attenuates renal fibrosis by suppressing the activation of TGF-beta1/Smad3 signaling pathway in HK-2 cells. Thus, RACK1 may serve as a novel regulator of renal fibrosis. PMID: 29039466
MSP analysis from 81 Acute coronary syndrome (ACS)samples, 74 SCAD samples and 53 healthy samples, and Sequenom MassARRAY analysis, confirmed that differential CpG methylation of SMAD3 was significantly corrected with the reference results of the HumanMethylation450 array. PMID: 29115576
Smad3 knockdown could restore the inhibition of cell proliferation induced by FSTL1 overexpression in MDAMB231FSTL1 cells, indicating that the antiproliferative effect of FSTL1 overexpression may be associated with Smad3 involved TGFbeta signaling pathway regulation. This study identified FSTL1 as an inhibitor of cell proliferation in MDAMB231 and 231BR cell lines PMID: 29048681
miR-195 inhibited proliferation and induced apoptosis of vascular smooth muscle cells, which was abated by Smad3 overexpression PMID: 28665537
SMAD3 SNP rs422342 is statistically associated with intervertebral disc degeneration in Greek population. PMID: 28662992
we observed that SMAD3 rs1065080 single nucleotide gene polymorphisms were significantly associated with patient susceptibility to intracranial arterial aneurysms PMID: 28988651
Smad3 binds with type I TGF-beta receptor (TRI) even in unstimulated cells. PMID: 27641076
This study demonstrates that Smad3 protein had low expression in ACTH-Pituitary Adenoma Development. PMID: 29524699
data suggest that TGF-beta stimulated the expression of ChPF and sGAG synthesis in nucleus pulposus cells through Smad3, RhoA/ROCK1 and the three MAPK signaling pathways. PMID: 28608941
These results suggested that FXR may serve as an important negative regulator for manipulating Smad3 expression, and the FXR/Smad3 pathway may be a novel target for the treatment of renal fibros PMID: 27853248
SMad3 role in TGF-beta/SMAD pathway signal transduction PMID: 28320972
ERK1/2 mediates Heme oxygenase-1 or CO-induced Smad3 phosphorylation at Thr179. PMID: 29524413
Participants' data and peripheral blood samples were collected, and three Smad3 CpG loci were examined. Smad3 mRNA expression was significantly higher in the patient group than in the negative control group but did not differ between the two control groups. PMID: 28562330
The critical roles of miR-16-5p-Smad3 pathway in melatonin-induced growth defects of gastric cancers. PMID: 29359963
TGFbeta1 signalling is associated with activation of SMAD3 at the ciliary base. PMID: 27748449
exaggerated WNT-5B expression upon cigarette smoke exposure in the bronchial epithelium of COPD patients leads to TGF-beta/Smad3-dependent expression of genes related to airway remodelling PMID: 27126693
HSF1 activity is decreased in fibrotic hearts. HSF1 inhibits phosphorylation and nuclear distribution of Smad3 via direct binding to Smad3. Active Smad3 blocks the anti-fibrotic effect of HSF1. PMID: 28091697
miR-142-5p plays as a negative regulator in TGF-beta pathway by targeting SMAD3 and suppresses TGF-beta-induced growth inhibition in cancer cells. PMID: 27683030
Authors were able to confirm expression of SMAD3 in intact and degraded cartilage of the knee and hip. Our findings provide the first systematic evaluation of pleiotropy between OA and BMD, highlight genes with biological relevance to both traits, and establish a robust new OA genetic risk locus at SMAD3. PMID: 28934396
A bioinformatics analysis and luciferase reporter assay identified Smad3 as a direct target gene of miR-216b, and Smad3 expression was reduced by miR-216b overexpression at both the mRNA and protein levels. PMID: 28356485
Because the expression of these genes correlates with cell shape, these are likely mechanosensitive genes that regulate SMAD3 and/or RELA activation in response to mechanical cues PMID: 27864353
SMAD3 transcription facttor binds RNA with large internal loops or bulges with high apparent affinity, suggesting a biological role for RNA binding by SMAD3. PMID: 29036649
Case Report: internal mammary artery aneurysms in sisters with SMAD3 mutation. PMID: 28286188
High Smad3 expression is associated with invasion and metastasis in pancreatic ductal adenocarcinoma. PMID: 26908446
New evidence suggests that SMAD3 activation may serve as a critical converging point of dysregulated TGFB superfamily signaling and genetic aberrations in human granulosa cell tumor development (review). PMID: 27683263
We find that DIGIT is divergent to Goosecoid (GSC) and expressed during endoderm differentiation. Deletion of the SMAD3-occupied enhancer proximal to DIGIT inhibits DIGIT and GSC expression and definitive endoderm differentiation. PMID: 27705785
ANP inhibits TGF-beta1-induced EMT in 16HBE-14o and A549 cells through cGMP/PKG signaling, by which it targets TGF-beta1/Smad3 via attenuating phosphorylation of Smad3. These findings suggest the potential of ANP in the treatment on pulmonary diseases with airway remodeling. PMID: 28229930
Sec8 regulates N-cadherin expression by controlling Smad3 and Smad4 expression through CBP, thereby mediating the epithelial-mesenchymal transition. PMID: 27769780
Particularly, galangin effectively inhibits phosphorylation of the Thr-179 site at Smad3 linker region through suppression of CDK4 phosphorylation. Thus, galangin can be a promising candidate as a selective inhibitor to suppress phosphorylation of Smad3 linker region. PMID: 29097203
Up-regulation of miR-195 suppressed cell migration and invasion in vitro. Smad3 was verified as a direct target of miR-195, which was further confirmed by the inverse expression of miR-195 and Smad3 in patients' specimens. PMID: 27206216
In human primary tubular epithelial cells, inhibition of HIF sensing prolylhydroxylases by DMOG or exposure of the cells to hypoxia upregulated Smad3 expression and enhanced its translocation to the nucleus. PMID: 27155083
Findings demonstrate that TGFbeta1 allows tumors to evade host immune responses in part through enhanced SMAD3-mediated PD-1 expression on tumor infiltrating lymphocytes. PMID: 27683557
Store-operated calcium entry via Orai1 in mesangial cells negatively regulates the TGF-beta1/Smad3 signaling pathway. PMID: 28637791
TF-induced microvessel stabilization is regulated via PAR2-SMAD3 that is indispensable for the maintenance of vascular integrity. PMID: 26658897
stablish PPM1A as a novel repressor of the SMAD3 pathway in renal fibrosis PMID: 27328942
Methylation in SMAD3 was selectively increased in asthmatic children of asthmatic mothers and was associated with childhood asthma risk PMID: 28011059
a direct crosstalk between the STAT3 and Smad3 signaling pathways that may contribute to tumor development and inflammation. PMID: 26616859
It is reported here that TGF-beta directly regulates alternative splicing of cancer stem cell marker CD44 through a phosphorylated threonine179 of SMAD3-mediated interaction with RNA-binding protein PCBP1. PMID: 27746021
Bcl-3 knockdown enhanced the degradation of Smad3 but not Smad2 following TGFbeta treatment. PMID: 27906182

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Proteins are sensitive to heat, and freeze-drying can preserve the activity of the majority of proteins. It improves protein stability, extends storage time, and reduces shipping costs. However, freeze-drying can also lead to the loss of the active portion of the protein and cause aggregation and denaturation issues. Nonetheless, these adverse effects can be minimized by incorporating protective agents such as stabilizers, additives, and excipients, and by carefully controlling various lyophilization conditions.

Commonly used protectant include saccharides, polyols, polymers, surfactants, some proteins and amino acids etc. We usually add 8% (mass ratio by volume) of trehalose and mannitol as lyoprotectant. Trehalose can significantly prevent the alter of the protein secondary structure, the extension and aggregation of proteins during freeze-drying process; mannitol is also a universal applied protectant and fillers, which can reduce the aggregation of certain proteins after lyophilization.

Our protein products do not contain carrier protein or other additives (such as bovine serum albumin (BSA), human serum albumin (HSA) and sucrose, etc., and when lyophilized with the solution with the lowest salt content, they often cannot form A white grid structure, but a small amount of protein is deposited in the tube during the freeze-drying process, forming a thin or invisible transparent protein layer.

Reminder: Before opening the tube cap, we recommend that you quickly centrifuge for 20-30 seconds in a small centrifuge, so that the protein attached to the tube cap or the tube wall can be aggregated at the bottom of the tube. Our quality control procedures ensure that each tube contains the correct amount of protein, and although sometimes you can't see the protein powder, the amount of protein in the tube is still very precise.

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