Phospho-Smad1/5/9 (Ser463+Ser465) Antibody - #AF8313

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製品: Phospho-Smad1/5/9 (Ser463+Ser465) Antibody
カタログ: AF8313
タンパク質の説明: Rabbit polyclonal antibody to Phospho-Smad1/5/9 (Ser463+Ser465)
アプリケーション: WB IHC
Cited expt.: WB
反応性: Human, Rat, Monkey
予測: Pig, Zebrafish, Bovine, Sheep, Rabbit, Dog, Chicken, Xenopus
分子量: 56kDa; 52kD(Calculated).
ユニプロット: Q15797 | Q99717 | O15198
RRID: AB_2840375

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製品説明

ソース:
Rabbit
アプリケーション:
WB 1:1000-3000, IHC 1:50-1:200
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

反応性:
Human,Rat,Monkey
予測:
Pig(100%), Zebrafish(100%), Bovine(100%), Sheep(100%), Rabbit(100%), Dog(100%), Chicken(100%), Xenopus(100%)
クローナリティ:
Polyclonal
特異性:
Phospho-Smad1/5/9 (Ser463+Ser465) Antibody detects endogenous levels of Smad1/5/9 only when phosphorylated at Ser463+Ser465.
RRID:
AB_2840375
引用形式: Affinity Biosciences Cat# AF8313, RRID:AB_2840375.
コンジュゲート:
Unconjugated.
精製:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
保存:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
別名:

折りたたみ／展開

BSP-1; BSP1; HsMAD1; JV4-1; JV41; MAD homolog 1; MAD mothers against decapentaplegic homolog 1; Mad related protein 1; Mad-related protein 1; MADH1; MADR1; Mothers against decapentaplegic homolog 1; Mothers against DPP homolog 1; SMA- AND MAD-RELATED PROTEIN 1; SMAD 1; SMAD family member 1; SMAD mothers against DPP homolog 1; Smad1; SMAD1_HUMAN; TGF beta signaling protein 1; Transforming growth factor-beta-signaling protein 1; DKFZp781C1895; DKFZp781O1323; Dwfc; hSmad5; JV5 1; JV5-1; MAD homolog 5; MAD, mothers against decapentaplegic homolog 5; MADH 5; MADH5; Mothers against decapentaplegic homolog 5; mothers against decapentaplegic, drosophila, homolog of, 5; Mothers against DPP homolog 5; MusMLP; SMA and MAD related protein 5; SMAD 5; SMAD family member 5; SMAD, mothers against DPP homolog 5; Smad5; SMAD5_HUMAN; MAD homolog 9; Madh6; Mothers against decapentaplegic; Mothers against decapentaplegic homolog 9; Mothers against DPP homolog 9; SMAD 9; SMAD family member 9; SMAD, mothers against DPP homolog 9 (Drosophila); SMAD8A; SMAD8B; Smad9; SMAD9_HUMAN;

免疫原

免疫原:

A synthesized peptide derived from human Smad1/5/9 around the phosphorylation site of Ser463+Ser465.

Uniprot:

Q15797(SMAD1_HUMAN) >>Visit HPA database.

Q99717(SMAD5_HUMAN) >>Visit HPA database.

O15198(SMAD9_HUMAN) >>Visit HPA database.

遺伝子(ID):
SMAD5(4090) | SMAD1(4086) | SMAD9(4093)
発現特異性:
Q15797 SMAD1_HUMAN:

Ubiquitous. Highest expression seen in the heart and skeletal muscle.

Q99717 SMAD5_HUMAN:

Ubiquitous.

O15198 SMAD9_HUMAN:

Expressed in heart, brain, placenta, lung, skeletal muscle, prostate, testis, ovary and small intestine. Also expressed in fetal brain, lung and kidney.

タンパク質配列:
MNVTSLFSFTSPAVKRLLGWKQGDEEEKWAEKAVDALVKKLKKKKGAMEELEKALSCPGQPSNCVTIPRSLDGRLQVSHRKGLPHVIYCRVWRWPDLQSHHELKPLECCEFPFGSKQKEVCINPYHYKRVESPVLPPVLVPRHSEYNPQHSLLAQFRNLGQNEPHMPLNATFPDSFQQPNSHPFPHSPNSSYPNSPGSSSSTYPHSPTSSDPGSPFQMPADTPPPAYLPPEDPMTQDGSQPMDTNMMAPPLPSEINRGDVQAVAYEEPKHWCSIVYYELNNRVGEAFHASSTSVLVDGFTDPSNNKNRFCLGLLSNVNRNSTIENTRRHIGKGVHLYYVGGEVYAECLSDSSIFVQSRNCNYHHGFHPTTVCKIPSGCSLKIFNNQEFAQLLAQSVNHGFETVYELTKMCTIRMSFVKGWGAEYHRQDVTSTPCWIEIHLHGPLQWLDKVLTQMGSPHNPISSVS

MTSMASLFSFTSPAVKRLLGWKQGDEEEKWAEKAVDALVKKLKKKKGAMEELEKALSSPGQPSKCVTIPRSLDGRLQVSHRKGLPHVIYCRVWRWPDLQSHHELKPLDICEFPFGSKQKEVCINPYHYKRVESPVLPPVLVPRHNEFNPQHSLLVQFRNLSHNEPHMPQNATFPDSFHQPNNTPFPLSPNSPYPPSPASSTYPNSPASSGPGSPFQLPADTPPPAYMPPDDQMGQDNSQPMDTSNNMIPQIMPSISSRDVQPVAYEEPKHWCSIVYYELNNRVGEAFHASSTSVLVDGFTDPSNNKSRFCLGLLSNVNRNSTIENTRRHIGKGVHLYYVGGEVYAECLSDSSIFVQSRNCNFHHGFHPTTVCKIPSSCSLKIFNNQEFAQLLAQSVNHGFEAVYELTKMCTIRMSFVKGWGAEYHRQDVTSTPCWIEIHLHGPLQWLDKVLTQMGSPLNPISSVS

MHSTTPISSLFSFTSPAVKRLLGWKQGDEEEKWAEKAVDSLVKKLKKKKGAMDELERALSCPGQPSKCVTIPRSLDGRLQVSHRKGLPHVIYCRVWRWPDLQSHHELKPLECCEFPFGSKQKEVCINPYHYRRVETPVLPPVLVPRHSEYNPQLSLLAKFRSASLHSEPLMPHNATYPDSFQQPPCSALPPSPSHAFSQSPCTASYPHSPGSPSEPESPYQHSVDTPPLPYHATEASETQSGQPVDATADRHVVLSIPNGDFRPVCYEEPQHWCSVAYYELNNRVGETFQASSRSVLIDGFTDPSNNRNRFCLGLLSNVNRNSTIENTRRHIGKGVHLYYVGGEVYAECVSDSSIFVQSRNCNYQHGFHPATVCKIPSGCSLKVFNNQLFAQLLAQSVHHGFEVVYELTKMCTIRMSFVKGWGAEYHRQDVTSTPCWIEIHLHGPLQWLDKVLTQMGSPHNPISSVS

種類予測

種類予測:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Bovine
100
Sheep
100
Dog
100
Xenopus
100
Zebrafish
100
Chicken
100
Rabbit
100
Horse
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

研究背景

機能:

Transcriptional modulator activated by BMP (bone morphogenetic proteins) type 1 receptor kinase. SMAD1 is a receptor-regulated SMAD (R-SMAD). SMAD1/OAZ1/PSMB4 complex mediates the degradation of the CREBBP/EP300 repressor SNIP1. May act synergistically with SMAD4 and YY1 in bone morphogenetic protein (BMP)-mediated cardiac-specific gene expression.

PTMs:

Phosphorylation of the C-terminal SVS motif by BMP type 1 receptor kinase activates SMAD1 by promoting dissociation from the receptor and trimerization with SMAD4.

Ubiquitinated by SMAD-specific E3 ubiquitin ligase SMURF1, leading to its degradation. Monoubiquitinated, leading to prevent DNA-binding. Deubiquitination by USP15 alleviates inhibition and promotes activation of TGF-beta target genes. Dephosphorylation, probably by PPM1A, induces its export from the nucleus to the cytoplasm (By similarity).

細胞の位置付け:

Cytoplasm. Nucleus.
Note: Cytoplasmic in the absence of ligand. Migrates to the nucleus when complexed with SMAD4 (PubMed:15647271). Co-localizes with LEMD3 at the nucleus inner membrane (PubMed:15647271). Exported from the nucleus to the cytoplasm when dephosphorylated (By similarity).

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
組織特異性:

Ubiquitous. Highest expression seen in the heart and skeletal muscle.

タンパク質ファミリー:

The MH2 domain mediates phosphorylation-dependent trimerization through L3 loop binding of phosphoserines in the adjacent subunit.

Belongs to the dwarfin/SMAD family.

機能:

Transcriptional modulator activated by BMP (bone morphogenetic proteins) type 1 receptor kinase. SMAD5 is a receptor-regulated SMAD (R-SMAD).

PTMs:

Phosphorylated on serine by BMP (bone morphogenetic proteins) type 1 receptor kinase.

Ubiquitin-mediated proteolysis by SMAD-specific E3 ubiquitin ligase SMURF1.

細胞の位置付け:

Cytoplasm. Nucleus.
Note: Cytoplasmic in the absence of ligand. Migrates to the nucleus when complexed with SMAD4.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
組織特異性:

Ubiquitous.

タンパク質ファミリー:

Belongs to the dwarfin/SMAD family.

機能:

Transcriptional modulator activated by BMP (bone morphogenetic proteins) type 1 receptor kinase. SMAD9 is a receptor-regulated SMAD (R-SMAD).

PTMs:

Phosphorylated on serine by BMP (bone morphogenetic proteins) type 1 receptor kinase.

細胞の位置付け:

Cytoplasm. Nucleus.
Note: In the cytoplasm in the absence of ligand. Migration to the nucleus when complexed with SMAD4 (By similarity).

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
組織特異性:

Expressed in heart, brain, placenta, lung, skeletal muscle, prostate, testis, ovary and small intestine. Also expressed in fetal brain, lung and kidney.

タンパク質ファミリー:

Belongs to the dwarfin/SMAD family.

研究領域

· Cellular Processes > Cellular community - eukaryotes > Signaling pathways regulating pluripotency of stem cells.   (View pathway)

· Environmental Information Processing > Signal transduction > TGF-beta signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > Hippo signaling pathway.   (View pathway)

· Human Diseases > Cancers: Overview > Transcriptional misregulation in cancer.

参考文献

1). Magnesium-enriched microenvironment promotes odontogenic differentiation in human dental pulp stem cells by activating ERK/BMP2/Smads signaling. Stem Cell Research & Therapy, 2019 (PubMed: 31823825) [IF=7.5]

Application: WB    Species: Human    Sample: DPSCs

Fig. 6 The ERK and BMP2 signaling pathway is activated by high extracellular Mg2+ in DPSCs during odontogenic differentiation. a ERK phosphorylation was significantly enhanced in DPSCs treated with 1 mM, 5 mM, and 10 mM Mg2+ compared with the 0 mM Mg2+ group, but p38 and JNK phosphorylation amounts were unchanged. b ERK phosphorylation was reduced by 2-APB. c Consistently, the protein levels of BMP2, BMPR1, and phosphorylated Smad1/5/9 were significantly increased in DPSCs exposed to high extracellular Mg2+. d BMP2, BMPR1, and phosphorylated Smad1/5/9 protein amounts were decreased by 2-APB
2). Maternal supplementation with mulberry-leaf flavonoids improves the development of skeletal muscle in the offspring of chickens. Animal nutrition (Zhongguo xu mu shou yi xue hui), 2024 (PubMed: 39035983) [IF=6.1]
3). BMP8B Activates Both SMAD2/3 and NF-κB Signals to Inhibit the Differentiation of 3T3-L1 Preadipocytes into Mature Adipocytes. Nutrients, 2023 (PubMed: 38201894) [IF=5.9]
4). Bmp8a deletion leads to obesity through regulation of lipid metabolism and adipocyte differentiation. Communications Biology, 2023 (PubMed: 37553521) [IF=5.9]

Application: WB    Species: Mouse    Sample: 3T3-L1 cells

Fig. 5 Bmp8a activates Smad2/3 signaling to inhibit adipocyte differentiation in 3T3-L1 cells. a–d Representative western blot analysis and quantification of changes in p-Smad1/5/8, p-Smad2/3, p-ERK1/2, p-p38 MAPK, and p-JNK expression in LV-bmp8a cells (a, b) or LV-Bmp8a cells (c, d). Protein expression levels were quantified using ImageJ software and normalized to the amount of total protein (n = 3). e, f Representative Oil Red O staining photographs of LV-bmp8a and LV-Bmp8a 3T3-L1 cells were induced to adipogenic in the presence of DMH1 or TP0427736 HCL, dimethylsulfoxide (DMSO) as a vehicle and subjected to OD492 quantifications (n = 3). Scale bar = 20 µm. g Schematic diagram of BMP8 mediated signal transduction. BMP8 can activate Smad1/5/8 signal transduction through the receptor complex formed by type I receptor ALK2, ALK3, or ALK6 and type II receptor ACVR2A or BMPR2. Meanwhile, BMP8 can also activate Smad2/3 signal transduction through the receptor complex formed by type I receptors ALK4 or ALK5 and type II receptors ACVR2A, ACVR2B, or TGFBR2. h Non-expression of mouse Alk6 gene in 3T3-L1 cells (n = 3). i The qPCR quantification of the type I receptor (Alk2, Alk3, Alk4, Alk5, Alk7) and type II receptor (Acvr2a, Acvr2b, Bmpr2, Tgrβr2) transcripts expressed in 3T3-L1 cells (n = 3). j, k Quantification of the activity of BRE-driven luciferase reporters with pCMV-bmp8a (j) or pCMV-Bmp8a (k) cotransfected with pCMV-Alk2, pCMV-Alk3, pCMV-Bmpr2, pCMV-Acrv2a, respectively (n = 3). Renilla luciferase was used as the internal control. l, m Quantification of the activity of CAGA-driven luciferase reporters with pCMV-bmp8a (l) or pCMV-Bmp8a (m) cotransfected with pCMV-Alk2, pCMV-Alk3, pCMV-Bmpr2, and pCMV-Acrv2a, respectively (n = 3). Renilla luciferase was used as the internal control. Data were representative of at least three independent experiments. Data were analyzed by One-way ANOVA and presented as mean ± SD
5). BMP8B Activates Both SMAD2/3 and NF-κB Signals to Inhibit the Differentiation of 3T3-L1 Preadipocytes into Mature Adipocytes. Nutrients, 2023 (PubMed: 38201894) [IF=5.9]

Application: WB    Species: Mouse    Sample:

Figure 3 BMP8B triggers SMAD2/3 signaling to suppress adipogenesis. (A,B) Analysis using immunoblotting and quantification was conducted to assess the protein levels of p-SMAD1/5/8, p-SMAD2/3, p-ERK1/2, p-p38 MAPK, and p-JNK in LV-Bmp8b. (C) A model of BMPs-associated signal transduction. (D) Quantification was performed to determine the luciferase reporter activity driven by BRE, which pCMV-Bmp8b cotransfected with pCMV-Alk2, pCMV-Alk3, pCMV-Bmpr2, pCMV-Acrv2a, respectively. (E) Quantification was performed to determine the luciferase reporter activity driven by CAGA, which pCMV-Bmp8b cotransfected with pCMV-Alk2, pCMV-Alk4, pCMV-Alk5, pCMV-Alk7, pCMV-Tgfβr2, pCMV-Acrv2a, and pCMV-Acrv2b, respectively. (F,G) In the presence of DMH1 or TP0427736 HCL, the cells were induced to differentiate into adipocytes. On Day 8, Oil Red O staining was performed (F). Quantification of lipid content after adipogenic differentiation (G). Scale bar = 20 µm. The symbols in the charts represent three biological replicates. The data were presented as mean ± SD and analyzed using one-way ANOVA (ns not significant, ** p < 0.01, *** p < 0.001).
6). Alternative splicing of EZH2 regulated by SNRPB mediates hepatocellular carcinoma progression via BMP2 signaling pathway. iScience, 2024 (PubMed: 39850359) [IF=5.8]
7). PTEN Reduces BMP9-Induced Osteogenic Differentiation Through Inhibiting Wnt10b in Mesenchymal Stem Cells. Frontiers in cell and developmental biology, 2021 (PubMed: 33614622) [IF=4.6]

Application: WB    Species: Mouse    Sample: C3H10T1/2 cells

Figure 6 Effects of CREB and BMP/Smad signaling on the expression of Wnt10b in C3H10T1/2 cells. (A) Western blot assay shows the effect of PTEN and/or BMP9 on CREB and phospho-CREB (p-CREB) in C3H10T1/2 cells. (B) Western blot assay shows the effect of PTEN knockdown and/or BMP9 on CREB and p-CREB in C3H10T1/2 cells. (C) Quantification of Western blot assay shows the effect of PTEN and/or BMP9 on p-CREB in C3H10T1/2 cells (**p < 0.01 vs. control; #p < 0.05 and ##p < 0.01 vs. groups treated with BMP9 only). (D) Quantification of Western blot assay shows effect of PTEN knockdown and/or BMP9 on p-CREB in C3H10T1/2 cells (**p < 0.01 vs. control; #p < 0.05 and ##p < 0.01 vs. groups treated with BMP9 only). (E) Western blot assay shows the effect of PTEN and/or BMP9 on Smad1/5/9 and p-Smad1/5/9 in C3H10T1/2 cells. (F) Western blot assay shows the effect of PTEN knockdown and/or BMP9 on Smad1/5/9 and p-Smad1/5/9 in C3H10T1/2 cells. (G) Quantification of Western blot assay shows the effect of PTEN and/or BMP9 on p-Smad1/5/9 in C3H10T1/2 cells (**p < 0.01 vs. control; ##p < 0.01 vs. groups treated with BMP9 only). (H) Quantification of Western blot assay shows effect of PTEN knockdown and/or BMP9 on p-Smad1/5/9 in C3H10T1/2 cells (**p < 0.01 vs. control; ##p < 0.01 vs. groups treated with BMP9 only). (I) IP assay shows the interaction between p-Smad1/5/9 and p-CREB in C3H10T1/2 cells. (J) IP assay shows the interaction between p-CREB and p-Smad1/5/9 in C3H10T1/2 cells. (K) ChIP assay shows the enrichment of p-CREB or p-Smad1/5/9 in the promoter region of Wnt10b C3H10T1/2 cells.
8). Wnt/β-Catenin Promotes the Osteoblastic Potential of BMP9 Through Down-Regulating Cyp26b1 in Mesenchymal Stem Cells. Tissue Engineering and Regenerative Medicine, 2023 (PubMed: 37010733) [IF=4.4]
9). Kartogenin Improves Osteogenesis of Bone Marrow Mesenchymal Stem Cells via Autophagy. Stem Cells International, 2022 (PubMed: 36591373) [IF=3.8]
10). Wnt and Smad signaling pathways synergistically regulated the osteogenic differentiation of fibroblasts in ankylosing spondylitis. TISSUE & CELL, 2022 (PubMed: 35753224) [IF=2.7]
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