产品: Smad4 抗体
货号: AF5247
描述: Rabbit polyclonal antibody to Smad4
应用: WB IHC IF/ICC
反应: Human, Mouse, Rat
预测: Pig, Bovine, Horse, Sheep, Rabbit, Dog
分子量: 65 kDa; 60kD(Calculated).
蛋白号: Q13485
RRID: AB_2837733

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 50ul RMB¥ 1250 现货
 100ul RMB¥ 2300 现货
 200ul RMB¥ 3000 现货

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产品描述

来源:
Rabbit
应用:
WB 1:500-1:2000, IF/ICC 1:100-1:500, IHC 1:50-1:200
*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:
Pig(100%), Bovine(100%), Horse(100%), Sheep(100%), Rabbit(100%), Dog(100%)
克隆:
Polyclonal
特异性:
Smad4 Antibody detects endogenous levels of total Smad4.
RRID:
AB_2837733
引用格式: Affinity Biosciences Cat# AF5247, RRID:AB_2837733.
偶联:
Unconjugated.
纯化:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
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

(Small) Mothers Against Decapentaplegic; Deleted in Pancreatic Carcinoma 4; Deleted in Pancreatic Carcinoma; Deleted in pancreatic carcinoma locus 4; Deletion target in pancreatic carcinoma 4; DPC 4; DPC4; hSMAD4; JIP; MAD homolog 4; MAD mothers against decapentaplegic Drosophila homolog 4; MAD mothers against decapentaplegic homolog 4; MADH 4; MADH4; Med; Medea; Mothers against decapentaplegic homolog 4; Mothers against decapentaplegic, Drosophila, homolog of, 4; Mothers against DPP homolog 4; MYHRS; OTTHUMP00000163548; SMA- and MAD-related protein 4; SMAD 4; SMAD family member 4; SMAD mothers against DPP homolog 4; SMAD4; SMAD4_HUMAN;

抗原和靶标

免疫原:
Uniprot:
基因/基因ID:
描述:
Common SMAD (co-SMAD) is the coactivator and mediator of signal transduction by TGF-beta (transforming growth factor). Component of the heterotrimeric SMAD2/SMAD3-SMAD4 complex that forms in the nucleus and is required for the TGF-mediated signaling.
序列:
MDNMSITNTPTSNDACLSIVHSLMCHRQGGESETFAKRAIESLVKKLKEKKDELDSLITAITTNGAHPSKCVTIQRTLDGRLQVAGRKGFPHVIYARLWRWPDLHKNELKHVKYCQYAFDLKCDSVCVNPYHYERVVSPGIDLSGLTLQSNAPSSMMVKDEYVHDFEGQPSLSTEGHSIQTIQHPPSNRASTETYSTPALLAPSESNATSTANFPNIPVASTSQPASILGGSHSEGLLQIASGPQPGQQQNGFTGQPATYHHNSTTTWTGSRTAPYTPNLPHHQNGHLQHHPPMPPHPGHYWPVHNELAFQPPISNHPAPEYWCSIAYFEMDVQVGETFKVPSSCPIVTVDGYVDPSGGDRFCLGQLSNVHRTEAIERARLHIGKGVQLECKGEGDVWVRCLSDHAVFVQSYYLDREAGRAPGDAVHKIYPSAYIKVFDLRQCHRQMQQQAATAQAAAAAQAAAVAGNIPGPGSVGGIAPAISLSAAAGIGVDDLRRLCILRMSFVKGWGPDYPRQSIKETPCWIEIHLHRALQLLDEVLHTMPIADPQPLD

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

翻译修饰 - Q13485 作为底物

Site PTM Type Enzyme
T9 Phosphorylation Q9UBE8 (NLK)
S22 Phosphorylation
S32 Phosphorylation
K37 Acetylation
K45 Acetylation
K45 Ubiquitination
K48 Acetylation
K70 Ubiquitination
T77 Phosphorylation Q15831 (STK11)
Y95 Phosphorylation
K106 Acetylation
K113 Sumoylation
K113 Ubiquitination
K122 Ubiquitination
S138 Phosphorylation Q9UBE8 (NLK)
K159 Sumoylation
S178 Phosphorylation
T265 Phosphorylation
T269 Phosphorylation
T273 Phosphorylation
T277 Phosphorylation P28482 (MAPK1) , P27361 (MAPK3)
S343 Phosphorylation Q14680 (MELK)
K385 Ubiquitination
K428 Acetylation
S504 Phosphorylation
K507 Acetylation
K507 Ubiquitination
Y513 Phosphorylation
K519 Ubiquitination

研究背景

功能:

In muscle physiology, plays a central role in the balance between atrophy and hypertrophy. When recruited by MSTN, promotes atrophy response via phosphorylated SMAD2/4. MSTN decrease causes SMAD4 release and subsequent recruitment by the BMP pathway to promote hypertrophy via phosphorylated SMAD1/5/8. Acts synergistically with SMAD1 and YY1 in bone morphogenetic protein (BMP)-mediated cardiac-specific gene expression. Binds to SMAD binding elements (SBEs) (5'-GTCT/AGAC-3') within BMP response element (BMPRE) of cardiac activating regions (By similarity). Common SMAD (co-SMAD) is the coactivator and mediator of signal transduction by TGF-beta (transforming growth factor). Component of the heterotrimeric SMAD2/SMAD3-SMAD4 complex that forms in the nucleus and is required for the TGF-mediated signaling. Promotes binding of the SMAD2/SMAD4/FAST-1 complex to DNA and provides an activation function required for SMAD1 or SMAD2 to stimulate transcription. Component of the multimeric SMAD3/SMAD4/JUN/FOS complex which forms at the AP1 promoter site; required for synergistic transcriptional activity in response to TGF-beta. May act as a tumor suppressor. 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 by PDPK1.

Monoubiquitinated on Lys-519 by E3 ubiquitin-protein ligase TRIM33. Monoubiquitination hampers its ability to form a stable complex with activated SMAD2/3 resulting in inhibition of TGF-beta/BMP signaling cascade. Deubiquitination by USP9X restores its competence to mediate TGF-beta signaling.

细胞定位:

Cytoplasm. Nucleus.
Note: Cytoplasmic in the absence of ligand. Migrates to the nucleus when complexed with R-SMAD (PubMed:15799969). PDPK1 prevents its nuclear translocation in response to TGF-beta (PubMed:17327236).

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
亚基结构:

Found in a complex with SMAD1 and YY1 (By similarity). Interacts with CITED2 (By similarity). Monomer; in the absence of TGF-beta activation. Heterodimer; on TGF-beta activation. Composed of two molecules of a C-terminally phosphorylated R-SMAD molecule, SMAD2 or SMAD3, and one molecule of SMAD4 to form the transcriptional active SMAD2/SMAD3-SMAD4 complex. Found in a ternary complex composed of SMAD4, STK11/LKB1 and STK11IP. Interacts with ATF2, COPS5, DACH1, MSG1, SKI, STK11/LKB1, STK11IP and TRIM33. Interacts with ZNF423; the interaction takes place in response to BMP2 leading to activation of transcription of BMP target genes. Interacts with ZNF521; the interaction takes place in response to BMP2 leading to activation of transcription of BMP target genes. Interacts with USP9X. Interacts (via the MH1 and MH2 domains) with RBPMS. Interacts with WWTR1 (via coiled-coil domain). Component of the multimeric complex SMAD3/SMAD4/JUN/FOS which forms at the AP1 promoter site; required for synergistic transcriptional activity in response to TGF-beta. Interacts with CITED1. Interacts with PDPK1 (via PH domain) (By similarity). Interacts with VPS39; this interaction affects heterodimer formation with SMAD3, but not with SMAD2, and leads to inhibition of SMAD3-dependent transcription activation. Interactions with VPS39 and SMAD2 may be mutually exclusive. Interacts with ZC3H3 (By similarity). Interacts (via MH2 domain) with ZNF451 (via N-terminal zinc-finger domains). Identified in a complex that contains at least ZNF451, SMAD2, SMAD3 and SMAD4. Interacts weakly with ZNF8. Interacts with NUP93 and IPO7; translocates SMAD4 to the nucleus through the NPC upon BMP7 stimulation resulting in activation of SMAD4 signaling. Interacts with CREB3L1, the interaction takes place upon TGFB1 induction and SMAD4 acts as CREB3L1 coactivator to induce the expression of genes involved in the assembly of collagen extracellular matrix. Interacts with DLX1. Interacts with ZBTB7A; the interaction is direct and stimulated by TGFB1. Interacts with CREBBP; the recruitment of this transcriptional coactivator is negatively regulated by ZBTB7A. Interacts with EP300; the interaction with this transcriptional coactivator is negatively regulated by ZBTB7A. Interacts with HDAC1. Interacts (via MH2 domain) with ZMIZ1 (via SP-RING-type domain); in the TGF-beta signaling pathway increases the activity of the SMAD3/SMAD4 transcriptional complex.

蛋白家族:

The MH1 domain is required for DNA binding.

The MH2 domain is required for both homomeric and heteromeric interactions and for transcriptional regulation. Sufficient for nuclear import.

Belongs to the dwarfin/SMAD family.

研究领域

· Cellular Processes > Cell growth and death > Cell cycle.   (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 > Wnt 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: Viral > Hepatitis B.

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

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

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

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

· Human Diseases > Cancers: Specific types > Chronic myeloid leukemia.   (View pathway)

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

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

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

文献引用

1). Dietary protein levels changed the hardness of muscle by acting on muscle fiber growth and the metabolism of collagen in sub-adult grass carp (Ctenopharyngodon idella). Journal of Animal Science and Biotechnology, 2022 (PubMed: 36002862) [IF=7.0]

Application: WB    Species: grass carp    Sample: muscle

Fig. 3 Western blot analysis of relative protein expression in the muscle of grass carp. A Collagen transcription related factors; B Related factors regulating collagen mRNA translation; C Collagen degradation related factors. Values are means ± SD and n = 6 for each group. Different letters are significantly different (P < 0.05)

2). LINC00909 up-regulates pluripotency factors and promotes cancer stemness and metastasis in pancreatic ductal adenocarcinoma by targeting SMAD4. Biology direct, 2024 (PubMed: 38504385) [IF=5.5]

Application: WB    Species: Human    Sample: PDAC cells

Fig. 5. LINC00909 enhances the stemness of pancreatic ductal adenocarcinoma (PDAC) by inhibiting SMAD4 expression. (A–C) RT-qPCR analysis was performed to detect the mRNA levels of SMAD4 in LINC00909-overexpressing cells (A) and LINC00909-knockdown cells (B, C). (D–F) Western blotting analysis was conducted to examine the protein expression of SMAD4 in PDAC cells. (G, H) The efficiency of SMAD4 knockdown in PANC-1 (G) and AsPC-1 cells (H) was detected by RT-qPCR. (I, J) Representative images of sphere-formation assays in PANC-1/KD and AsPC-1/KD cells transfected with siSMAD4-2. (K, L) The protein levels of SMAD4, KLF4, c-Myc, JNK, p-JNK, JUN and p-JUN in PANC-1/LINC00909-KD (K) and AsPC-1/LINC00909-KD cells (L) transfected with siSMAD4-2 were analyzed by western blotting. (M) PANC-1 cells transfected with EV or vector encoding LINC00909 were treated with actinomycin D (1 mg/ml) at the indicated time point. (N, O) PANC-1 or AsPC-1 cells transfected with siNC, siLINC00909-3, or siLINC00909-6 were treated with actinomycin D (1 mg/ml) at the indicated time point. Total RNA was extracted and analyzed by RT-qPCR to examine the relative levels of SMAD4 mRNA (M–O). All *P 

3). Jiawei Maxing Shigan Tang alleviates radiation-induced lung injury via TGF-β1/Smad signaling pathway mediated by regulatory T cells. Journal of ethnopharmacology, 2024 (PubMed: 37944875) [IF=5.4]

Application: WB    Species: Human    Sample:

Fig. 1. Evaluation of the effects of CD4+CD25+ Tregs on TGF-β1/Smad pathway and the EMT process. (A) The release of TGF-β1 from Tregs was detected by ELISA. (B–F) Protein levels of Smad4, Vimentin, α-SMA, and E-cadherin in type II AECs co-cultured with Tregs supernatant were detected by Western blot. *P<0.05, **P<0.01, ***P<0.001, n = 3.

4). Comprehensive upstream and downstream regulatory analyses identify miR-675-3p as a potential prognostic biomarker in melanoma. Human Cell, 2021 (PubMed: 33400243) [IF=4.3]

Application: WB    Species: Human    Sample: melanoma A375 cells

Fig. 7 miR-675-3p may promote cell proliferation, regulate the cell cycle, and activate the TGF-β, HIF-1 signaling pathways. a miR- 675-3p facilitated the G0/G1–G2/M transition in human melanoma A375 cells, b miR-675-3p promoted cell proliferation. c The TGFβ2, Smad2/3, Smad4, and HIF1A protein levels in the TGF-β/SMAD and HIF-1 signaling pathways were signifcantly higher, while TGF β1 was decreased relative to the negative control group

5). Effects of dietary methionine on growth performance, muscle nutritive deposition, muscle fibre growth and type I collagen synthesis of on-growing grass carp (Ctenopharyngodon idella). The British journal of nutrition, 2021 (PubMed: 32718370) [IF=3.6]

Application: WB    Species: grass carp    Sample: muscle

Fig. 7. |Western blot analysis of protein expression of genes involved in type I collagen metabolism in the muscle of on-growing grass carp (Ctenopharyngodon idella) fed diets with graded levels of Met(g kg -1) for 60 days. Data represent means of three fish in each group, error bars indicate S.D. Values having different letters are significantly different

6). Astragaloside IV inhibits cell proliferation in vulvar squamous cell carcinoma through the TGF‐β/Smad signaling pathway. Dermatologic Therapy, 2019 (PubMed: 30536730) [IF=3.6]

Application: WB    Species: human    Sample: SW962 cells

Figure 3. |AS-Ⅳ reverses TGF-β/Smad signaling abnormalities and inhibits TGF-β1-mediated cell proliferation and anti-apoptosis in SW962 cells. SW962 cells were treated with TGF-β1 (10 ng/ml) alone or in combination with AS-Ⅳ (800 μg/ml) for 24 h and 48 h, respectively.Western blot assays evaluated the protein levels of TGF-βRI, TGF-βRII,Smad2/3, p-Smad2/3 and Smad4 (A,B).

7). Catalpol Alleviates Ang II-Induced Renal Injury Through NF‐κB Pathway and TGF-β1/Smads Pathway. Journal of Cardiovascular Pharmacology, 2022 (PubMed: 34654783) [IF=3.0]

8). Polysaccharides Extracted from Angelica sinensis (Oliv.) Diels Relieve the Malignant Characteristics of Glioma Cells through Regulating the MiR-373-3p-Mediated TGF-β/Smad4 Signaling Pathway. Evidence-based Complementary and Alternative Medicine, 2022 (PubMed: 35855826)

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