Phospho-Smad2 (Ser250) Antibody - #AF3450

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产品: 磷酸化 Smad2 (Ser250) 抗体
货号: AF3450
描述: Rabbit polyclonal antibody to Phospho-Smad2 (Ser250)
应用: WB IF/ICC
文献验证: WB
反应: Human, Mouse, Rat
预测: Zebrafish, Bovine, Horse, Sheep, Rabbit, Dog, Xenopus
分子量: 65kDa; 52kD(Calculated).
蛋白号: Q15796
RRID: AB_2834845

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产品描述

来源:
Rabbit
应用:
WB 1:500-1:2000, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: 适用于变性蛋白样本的免疫印迹检测. IHC: 适用于组织样本的石蜡(IHC-p)或冰冻(IHC-f)切片样本的免疫组化/荧光检测. IF/ICC: 适用于细胞样本的荧光检测. ELISA(peptide): 适用于抗原肽的ELISA检测.

反应:
Human, Mouse, Rat
预测:
Zebrafish(86%), Bovine(%), Horse(%), Sheep(%), Rabbit(%), Dog(%), Xenopus(%)
克隆:
Polyclonal
特异性:
Phospho-Smad2 (Ser250) Antibody detects endogenous levels of Smad2 only when phosphorylated at Serine 250.
RRID:
AB_2834845
引用格式: Affinity Biosciences Cat# AF3450, RRID:AB_2834845.
偶联:
Unconjugated.
纯化:
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.
别名:

展开/折叠

Drosophila, homolog of, MADR2; hMAD-2; HsMAD2; JV18; JV18-1; JV181; MAD; MAD homolog 2; MAD Related Protein 2; Mad-related protein 2; MADH2; MADR2; MGC22139; MGC34440; Mother against DPP homolog 2; Mothers against decapentaplegic homolog 2; Mothers against decapentaplegic, Drosophila, homolog of, 2; Mothers against DPP homolog 2; OTTHUMP00000163489; Sma and Mad related protein 2; Sma- and Mad-related protein 2 MAD; SMAD 2; SMAD family member 2; SMAD, mothers against DPP homolog 2; SMAD2; SMAD2_HUMAN;

抗原和靶标

免疫原:

A synthesized peptide derived from human Smad2 around the phosphorylation site of Ser250.

Uniprot:

Q15796(SMAD2_HUMAN) >>Visit HPA database.

基因/基因ID:
SMAD2(4087)
表达:
Q15796 SMAD2_HUMAN:

Expressed at high levels in skeletal muscle, endothelial cells, heart and placenta.

描述:
The protein encoded by this gene belongs to the SMAD, a family of proteins 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 mediates the signal of the transforming growth factor (TGF)-beta, and thus regulates multiple cellular processes, such as cell proliferation, apoptosis, and differentiation.
序列:
MSSILPFTPPVVKRLLGWKKSAGGSGGAGGGEQNGQEEKWCEKAVKSLVKKLKKTGRLDELEKAITTQNCNTKCVTIPSTCSEIWGLSTPNTIDQWDTTGLYSFSEQTRSLDGRLQVSHRKGLPHVIYCRLWRWPDLHSHHELKAIENCEYAFNLKKDEVCVNPYHYQRVETPVLPPVLVPRHTEILTELPPLDDYTHSIPENTNFPAGIEPQSNYIPETPPPGYISEDGETSDQQLNQSMDTGSPAELSPTTLSPVNHSLDLQPVTYSEPAFWCSIAYYELNQRVGETFHASQPSLTVDGFTDPSNSERFCLGLLSNVNRNATVEMTRRHIGRGVRLYYIGGEVFAECLSDSAIFVQSPNCNQRYGWHPATVCKIPPGCNLKIFNNQEFAALLAQSVNQGFEAVYQLTRMCTIRMSFVKGWGAEYRRQTVTSTPCWIELHLNGPLQWLDKVLTQMGSPSVRCSSMS

种属预测

种属预测:

score>80的预测可信度较高，可尝试用于WB检测。*预测模型主要基于免疫原序列比对，结果仅作参考，不作为质保凭据。

Species
Results
Score
Horse
100
Bovine
100
Sheep
100
Dog
100
Rabbit
100
Xenopus
86
Zebrafish
86
Pig
0
Chicken
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

研究背景

功能:

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 SMAD2/SMAD4 complex, activates transcription. May act as a tumor suppressor in colorectal carcinoma. Positively regulates PDPK1 kinase activity by stimulating its dissociation from the 14-3-3 protein YWHAQ which acts as a negative regulator.

翻译修饰:

Phosphorylated on one or several of Thr-220, Ser-245, Ser-250, and Ser-255. In response to TGF-beta, phosphorylated on Ser-465/467 by TGF-beta and activin type 1 receptor kinases. TGF-beta-induced Ser-465/467 phosphorylation declines progressively in a KMT5A-dependent manner. Able to interact with SMURF2 when phosphorylated on Ser-465/467, recruiting other proteins, such as SNON, for degradation. In response to decorin, the naturally occurring inhibitor of TGF-beta signaling, phosphorylated on Ser-240 by CaMK2. Phosphorylated by MAPK3 upon EGF stimulation; which increases transcriptional activity and stability, and is blocked by calmodulin. Phosphorylated by PDPK1.

In response to TGF-beta, ubiquitinated by NEDD4L; which promotes its degradation. Monoubiquitinated, leading to prevent DNA-binding (By similarity). Deubiquitination by USP15 alleviates inhibition and promotes activation of TGF-beta target genes. Ubiquitinated by RNF111, leading to its degradation: only SMAD2 proteins that are 'in use' are targeted by RNF111, RNF111 playing a key role in activating SMAD2 and regulating its turnover (By similarity).

Acetylated on Lys-19 by coactivators in response to TGF-beta signaling, which increases transcriptional activity. Isoform short: Acetylation increases DNA binding activity in vitro and enhances its association with target promoters in vivo. Acetylation in the nucleus by EP300 is enhanced by TGF-beta.

细胞定位:

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:9865696). On dephosphorylation by phosphatase PPM1A, released from the SMAD2/SMAD4 complex, and exported out of the nucleus by interaction with RANBP1 (PubMed:16751101, PubMed:19289081).

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
组织特异性:

Expressed at high levels in skeletal muscle, endothelial cells, heart and placenta.

蛋白家族:

Belongs to the dwarfin/SMAD family.

研究领域

· Cellular Processes > Cell growth and death > Cell cycle.   (View pathway)

· Cellular Processes > Transport and catabolism > Endocytosis.   (View pathway)

· Cellular Processes > Cell growth and death > Cellular senescence.   (View pathway)

· Cellular Processes > Cellular community - eukaryotes > Adherens junction.   (View pathway)

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

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

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

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

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

· Human Diseases > Infectious diseases: Parasitic > Chagas disease (American trypanosomiasis).

· Human Diseases > Infectious diseases: Viral > HTLV-I infection.

· Human Diseases > Cancers: Overview > Pathways in cancer.   (View pathway)

· Human Diseases > Cancers: Overview > Proteoglycans in cancer.

· Human Diseases > Cancers: Specific types > Colorectal cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Pancreatic cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Hepatocellular carcinoma.   (View pathway)

· Human Diseases > Cancers: Specific types > Gastric cancer.   (View pathway)

· Human Diseases > Immune diseases > Inflammatory bowel disease (IBD).

· Organismal Systems > Immune system > Th17 cell differentiation.   (View pathway)

· Organismal Systems > Endocrine system > Relaxin signaling pathway.

文献引用

1). Histone deacetylase 3 aberration inhibits Klotho transcription and promotes renal fibrosis. Cell death and differentiation, 2021 (PubMed: 33024274) [IF=13.7]

Application: WB    Species: Mouse    Sample: renal tissues

Fig. 6: Klotho derepression is essential for the antifibrosis function of HDAC3 inhibition.Mice receiving siRNA-Control (siCon) or siRNA-Klotho (siKL) were subgrouped into Sham, RGFP966, UUO, and RGFP966-treated UUO (n = 6). a Representative photomicrographs of kidney sections (Masson’s trichrome staining). The dark arrows indicate fibrotic areas. b Quantitation of renal fibrosis in a. The effects of siRNA-Klotho (P1), RGFP966 intervention (P2), and the interaction between siRNA-Klotho and RGFP966 intervention (P3) were indicated. c Western blots. The renal tissues from experimental mice in a were assayed for Klotho, α-SMA, BMP-7, collagen 1 (Colla 1), and d β-catenin (β-cat) and phosphorylated Smad2 (p-Smd2). Two samples from each group were shown. e Quantification of c and d. Data were presented as means ± SEM. *P 
2). Antifibrotic Effects of Tetrahedral Framework Nucleic Acids by Inhibiting Macrophage Polarization and Macrophage-myofibroblast Transition in Bladder Remodeling. Advanced Healthcare Materials, 2023 (PubMed: 36603196) [IF=10.0]
3). Methylation of BRD4 by PRMT1 regulates BRD4 phosphorylation and promotes ovarian cancer invasion. Cell death & disease, 2023 (PubMed: 37737256) [IF=8.1]

Application: WB    Species: Mouse    Sample: A2780 cells

Fig. 7 PRMT1-mediated BRD4 methylation is correlated with TGF-β. a and b IB analysis of TGF-β derived from SKOV3 cells and A2780 cells transfected with shPRMT1. c and d IB analysis of TGF-β derived from SKOV3 cells and A2780 cells treated with AMI-1 for 24, 48, and 72 h. e–g IB analysis of TGF-β derived from SKOV3 cells, A2780 cells, 293T cells stably expressed HA-BRD4-WT and HA-BRD4-R3K mutant. h and i IB analysis of TGF-β derived from BRD4 knockdown 293T cells and BRD4 knockdown A2780 cells stably expressed HA-BRD4-WT and HA-BRD4-R3K mutant. j IB analysis of p-Smad2 and p-Smad4 derived from A2780 cells transfected with shPRMT1-1 and shPRMT1-2. k IB analysis of TGF-β derived from A2780 cells infected with sgTGF-β. l CCK8 assays were performed in TGF-β knockout A2780 ovarian cancer cells for 24, 48, and 72 h. Statistical analysis of relative colony numbers was plotted. ****P 
4). Squalene epoxidase promotes hepatocellular carcinoma development by activating STRAP transcription and TGF-β/SMAD signalling. British Journal of Pharmacology, 2023 (PubMed: 36581319) [IF=6.8]
5). L-Glutamine alleviates osteoarthritis by regulating lncRNA-NKILA expression through the TGF-β1/SMAD2/3 signalling pathway. Clinical Science, 2022 (PubMed: 35730575) [IF=6.7]
6). Protective effect of remdesivir against pulmonary fibrosis in mice. Frontiers in Pharmacology, 2021 (PubMed: 34512328) [IF=5.6]

Application: WB    Species: Mice    Sample: lung tissues

FIGURE 10 Remdesivir suppress BLM-induced pulmonary fibrosis in mice via inhibiting TGF-β1-Smad and non-Smad signaling pathway in vivo. Protein levels of p-Smad2, Smad2, p-Smad3, Smad3, p-P38, P38, p-JNK, JNK, p-ERK, ERK, p-AKT, and AKT were verified by Western blot in lung tissues. GAPDH was used as an internal reference in densitometric analysis. Data was presented as the means ± SD, n = 3. ** p < 0.01, *** p < 0.001.
7). Myricetin ameliorates bleomycin-induced pulmonary fibrosis in mice by inhibiting TGF-β signaling via targeting HSP90β. BIOCHEMICAL PHARMACOLOGY, 2020 (PubMed: 32535102) [IF=5.3]
8). Guizhi Fuling pill attenuates liver fibrosis in vitro and in vivo via inhibiting TGF-β1/Smad2/3 and activating IFN-γ/Smad7 signaling pathways. Bioengineered, 2022 (PubMed: 35387552) [IF=4.2]

Application: WB    Species: human    Sample: LX-2 cells

Figure 2.| GZFL inhibits acetaldehyde-induced LX-2 cells activation through suppressing TGF-β1/Smad2/3 signaling and activating IFN-γ/STAT1/Smad7 signaling. LX-2 cells were exposed to acetaldehyde (AA; 400 μM) for 24 h, followed by exposure to colchicine (4 μg/ml) or GZFL (8 or 10 mg/ml) for another 24 h. (a, b) TGF-β1, TGF-βR2, CUGBP1, p-STAT1, p-Smad2, p-Smad3, Smad7,α-SMA and Collagen I expressions in LX-2 cells were detected by western blot assay.
9). 2-Hydroxypropyl-β-cyclodextrin Regulates the Epithelial to Mesenchymal Transition in Breast Cancer Cells by Modulating Cholesterol Homeostasis and Endoplasmic Reticulum Stress. Metabolites, 2021 (PubMed: 34436503) [IF=4.1]

Application: WB    Species: Human    Sample: MDA-MB-231 cells

Figure 5 The effect of low concentrations of HP-β-CD on ER stress, TGF-β/Smad signaling pathway, and the distribution of cholesterol in MDA-MB-231 cells. (A) MDA-MB-231 cells were treated with various lower concentration of HP-β-CD (0, 0.2, 0.4, 0.6, 0.8 mmol/L). Expression levels of E-cadherin and Vimentin. (B) Expression levels of GRP78 and CHOP. (C) Expression level of p-Smad2 and Smad2. (D) MDA-MB-231 cells were treated with various lower concentration of HP-β-CD (0, 0.6, 0.8 mmol/L). Expression levels of HMGCR and LXRα. (E) Co-staining of Filipin and immunofluorescence analyses for the content of cholesterol in ER. Magnification, ×400. The red fluorescence showed the ER. Scale bars represent 40μm. Data are presented as the mean ± SD (n = 3). * p < 0.05, ** p < 0.01 versus the control group.
10). Dapagliflozin inhibits ferroptosis and ameliorates renal fibrosis in diabetic C57BL/6J mice. Scientific reports, 2025 (PubMed: 40016517) [IF=3.8]
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