Invitrogen

Phospho-SMAD3 (Ser423, Ser425) Polyclonal Antibody

This Antibody was verified by Cell treatment to ensure that the antibody binds to the antigen stated. 查看细节

8篇引用文献

查看其他29个SMAD3抗体

Product Image Gallery

Phospho-SMAD3 (Ser423, Ser425) Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of SMAD3 (pSpS423/425) was done on 70% confluent log phase HeLa cells treated with TGF-beta (20 ng/mL for 15 min) cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with SMAD3 (pSpS423/425) Rabbit polyclonal Antibody (Product # 44-246G) at 1:250 dilution in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Fluor 488 Goat Anti-Rabbit IgG Secondary Antibody (Product # A-11008) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor 594 Phalloidin (Product # A12381). Panel d is a merged image showing nuclear translocation of SMAD3 (pSpS423/425). Panel e cytoplasmic localization. Panel f shows no primary antibody control. The images were captured at 20X magnification.

Phospho-SMAD3 (Ser423, Ser425) Antibody in Western Blot (WB) — Peptide Competition. Extracts of NMuMG cells unstimulated (1) or stimulated with 8 ng/mL TGF-beta for 20 minutes (2-5) were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. The membrane was blocked with a 5% BSA-TBST buffer for one hour at room temperature, then incubated with Smad3 (pSpS423/425) antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1, 2), the non-phosphorylated peptide corresponding to the phosphopeptide immunogen (3), a generic phosphoserine-containing peptide (4), or the phosphopeptide immunogen (5). After washing, the membrane was incubated with goat F (ab')2 anti-rabbit IgG HRP conjugate (Product # ALI4404) and signals were detected using the Pierce SuperSignal™ method. The data show that only the phosphopeptide corresponding to Smad3 (pSpS423/425) completely blocks the antibody signal, demonstrating the specificity of the antibody. The data also show upregulation of Smad3 (pSpS423/425) by stimulation with TGF-beta in this testing system. Note: The upper band is believed to be Smad3 in complex with Smad4.

Phospho-SMAD3 (Ser423, Ser425) Antibody — Detection of altered subcellular localization of the target protein by cell treatment demonstrates antibody specificity. Immunofluorescence analysis of Phospho-SMAD3 (Ser423, Ser425) using Phospho-SMAD3 (Ser423, Ser425) Rabbit Polyclonal Antibody (Product # 44-246G) shows translocation of SMAD3 from cytoplasm to nucleus of HeLa cells upon treatment with TGF-beta. {TM}

产品细节

Applications

Tested Dilution

Publications

免疫印迹 (WB)

1:1,000

View 5 publications 5 publications

免疫组化 (IHC)

-

View 2 publications 2 publications

免疫细胞化学 (ICC/IF)

1:250

View 1 publication 1 publication

产品规格

种属反应

Human, Mouse

已发表种属

Hamster, Human, Mouse, Non-human primate, Rat

宿主/亚型

Rabbit / IgG

分类

Polyclonal

类型

Antibody

抗原

The antiserum was produced against a chemically synthesized phosphopeptide derived from a region of human Smad3 that contains serine 423 and serine 425. This sequence is conserved in mouse.

偶联物

Unconjugated

形式

Liquid

纯化类型

Antigen affinity chromatography

保存液

Dulbecco's PBS, pH 7.3, with 1mg/mL BSA, 50% glycerol

内含物

0.05% sodium azide

保存条件

-20°C

RRID

AB_2533615

靶标信息

SMAD3 belongs to the SMAD family of proteins. They are 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.

仅用于科研。不用于诊断过程。未经明确授权不得转售。

生物信息学

蛋白别名: DKFZp586N0721; DKFZp686J10186; hMAD-3; hSMAD3; JV15-2; MAD homolog 3; mad homolog JV15-2; mad protein homolog; MAD, mothers against decapentaplegic homolog 3; MAD-3; mad3; MGC60396; mMad3; Mothers against decapentaplegic homolog 3; mothers against DPP homolog 3; SMA- and MAD-related protein 3; SMAD; SMAD 3; SMAD family member 3; SMAD, mothers against DPP homolog 3

基因别名: AU022421; HSPC193; HsT17436; JV15-2; LDS1C; LDS3; MADH3; SMAD3

UniProt ID: (Human) P84022, (Mouse) Q8BUN5

Entrez Gene ID: (Human) 4088, (Mouse) 17127

分子生物学功能: DNA-binding transcription factor gene-specific transcriptional regulator

negative regulation of transcription from RNA polymerase II promoter ureteric bud development response to hypoxia in utero embryonic development mesoderm formation somitogenesis liver development heart looping osteoblast development immune system development transcription, DNA-templated regulation of transcription from RNA polymerase II promoter transport activation of cysteine-type endopeptidase activity involved in apoptotic process immune response cell cycle arrest transforming growth factor beta receptor signaling pathway SMAD protein complex assembly endoderm development embryonic pattern specification positive regulation of gene expression positive regulation of alkaline phosphatase activity positive regulation of epithelial to mesenchymal transition regulation of striated muscle tissue development regulation of transforming growth factor beta receptor signaling pathway evasion or tolerance of host defenses by virus signal transduction involved in regulation of gene expression negative regulation of cell growth positive regulation of cell migration positive regulation of bone mineralization negative regulation of transforming growth factor beta receptor signaling pathway thyroid gland development primary miRNA processing positive regulation of chondrocyte differentiation positive regulation of interleukin-1 beta production regulation of transforming growth factor beta2 production positive regulation of transforming growth factor beta3 production activin receptor signaling pathway negative regulation of osteoblast proliferation nodal signaling pathway wound healing T cell activation positive regulation of transcription factor import into nucleus negative regulation of apoptotic process cell-cell junction organization negative regulation of fat cell differentiation negative regulation of osteoblast differentiation positive regulation of transcription, DNA-templated negative regulation of mitotic cell cycle positive regulation of transcription from RNA polymerase II promoter paraxial mesoderm morphogenesis developmental growth embryonic foregut morphogenesis embryonic cranial skeleton morphogenesis regulation of epithelial cell proliferation negative regulation of inflammatory response regulation of immune response positive regulation of positive chemotaxis regulation of binding positive regulation of stress fiber assembly positive regulation of focal adhesion assembly pericardium development transdifferentiation SMAD protein signal transduction negative regulation of wound healing lens fiber cell differentiation extrinsic apoptotic signaling pathway activation of cysteine-type endopeptidase activity involved in apoptotic signaling pathway positive regulation of extracellular matrix assembly skeletal system development osteoblast differentiation negative regulation of protein phosphorylation regulation of transcription, DNA-templated transcription from RNA polymerase II promoter gastrulation negative regulation of cell proliferation positive regulation of catenin import into nucleus intracellular signal transduction negative regulation of protein catabolic process protein stabilization positive regulation of canonical Wnt signaling pathway

参与通路

RNA polymerase II core promoter proximal region sequence-specific DNA binding transcription factor activity, RNA polymerase II core promoter sequence-specific core promoter proximal region sequence-specific DNA binding transcription factor activity, protein binding RNA polymerase II activating transcription factor binding transcription factor activity, sequence-specific DNA binding transforming growth factor beta receptor binding protein binding collagen binding transcription factor binding zinc ion binding protein kinase binding phosphatase binding transforming growth factor beta receptor, pathway-specific cytoplasmic mediator activity chromatin DNA binding ubiquitin protein ligase binding enhancer binding identical protein binding protein homodimerization activity ubiquitin binding bHLH transcription factor binding sequence-specific DNA binding transcription regulatory region DNA binding protein heterodimerization activity co-SMAD binding R-SMAD binding DNA binding chromatin binding double-stranded DNA binding beta-catenin binding enzyme binding SMAD binding metal ion binding