产品: 磷酸化 Smad4 (Thr276)[Thr277] 抗体
货号: AF8316
描述: Rabbit polyclonal antibody to Phospho-Smad4 (Thr276)[Thr277]
应用: WB IHC IF/ICC
反应: Human, Mouse, Rat
预测: Pig, Bovine, Horse, Sheep, Rabbit, Dog
分子量: 70kDa; 60kD(Calculated).
蛋白号: Q13485
RRID: AB_2840378

浏览相似产品>>

   规格 价格 库存
 100ul RMB¥ 2800 现货
 200ul RMB¥ 3800 现货

货期: 当天发货

联系销售

产品描述

来源:
Rabbit
应用:
WB 1:1000-3000, IHC 1:50-1:200, 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:
Pig(90%), Bovine(90%), Horse(100%), Sheep(100%), Rabbit(100%), Dog(90%)
克隆:
Polyclonal
特异性:
Phospho-Smad4 (Thr276) Antibody detects endogenous levels of Smad4 only when phosphorylated at Thr277, which site historically referenced as Thr276.
RRID:
AB_2840378
引用格式: Affinity Biosciences Cat# AF8316, RRID:AB_2840378.
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

(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:
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
Horse
100
Sheep
100
Rabbit
100
Pig
90
Bovine
90
Dog
90
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). Organoid modelling identifies that DACH1 functions as a tumour promoter in colorectal cancer by modulating BMP signalling. eBioMedicine (PubMed: 32512510) [IF=11.1]

Application: WB    Species: Human    Sample: HCT116 cells

Fig. 5. DACH1 promotes adenomas organoid formation via modulating BMP signalling. a DACH1 overexpression induced upregulation of cancer stem cell marker genes. b and c. Gene Ontology (GO) analysis showed that DACH1 overexpression induced the upregulation of stem cell signature genes and the downregulation of cell cycle arrest signature genes. d and e. DACH1 induced the downregulation of ATOH8, TGFb3, MSX1, NBL1, BMP7, and FKBP4 and the upregulation of FKBP8, ID1, and MAPK3 (experiments were performed in triplicate). f. IF images showing the colocalization of SMAD4 and DACH1 in the nuclei; scale bars=20 mm g-h. DACH1 coprecipitated endogenous SMAD4. Reverse immunoprecipitation was confirmed with an antiSMAD4 antibody. i. DACH1 overexpression increased the protein level of SMAD4 and decreased the level of phosphorylated SMAD4 (Thr276) [Thr277]. j and k. DACH1 knockdown led to an increase of the mRNA levels of NBL1 and BMP7 compared with the shNC group, and siRNA mediated SMAD4 knockdown in HCT116-shDACH1 cells eliminated the increase. l. DACH1 overexpression was sufficient to compensate for the withdrawal of Noggin and supported the formation of adenoma organoids. m. Addition of Noggin into the culture medium for 24 and 36 h decreased the mRNA levels of NBL1 and BMP7, which were increased by DACH1 knockdown in HCT116 cells. n and o. Overexpression of DACH1 upregulates LGR5, Notch1 and the protein level of NICD, while did not induce significant upregulation of HES1. Scale bars=20 mm. For 5d, 5e, 5k, 5 m and 5n, **P < 0.01, *P < 0.05, Student’s t-test. Error bars: mean§SD.

2). SPARCL1 promotes C2C12 cell differentiation via BMP7-mediated BMP/TGF-β cell signaling pathway. Cell Death & Disease (PubMed: 31699966) [IF=9.0]

Application: WB    Species: Mouse    Sample: C2C12 cells

Fig. 6 SPARCL1 regulates BMP7 expression and BMP/TGF-β cell signaling pathway. a, d shows the protein expression of BMP7 regulated by SPARCL1 activation and inhibition, respectively. C2C12 cells were induced to differentiate at 72 h. pSPgRNA-S-2 is the SPARCL1 activation group, while pSPgRNA is the blank control for SPARCL1 activation. NC is the negative control for SPARCL1 siRNA interference. b, c are greyscale scans of SPARCL1 and BMP7 proteins in A. e, f are grayscale scans of SPARCL1 and BMP7 proteins in D. i and p shows changes in the expression of BMP/ TGF-β-associated proteins when SPARCL1 was activated and inhibited, respectively, and C2C12 cells were induced to differentiate at 72 h. pSPgRNA-S-2 is the SPARCL1 activation group, while pSPgRNA is the blank control for SPARCL1 activation. NC is the negative control for SPARCL1 siRNA interference. j–o are greyscale scans of proteins in I. q–v are greyscale scans of proteins in P. **P values < 0.01 were considered as significant

3). TOPK mediates immune evasion of renal cell carcinoma via upregulating the expression of PD-L1. iScience (PubMed: 37404377) [IF=5.8]

Application: WB    Species: Human    Sample: 293T cells

Figure 6 TOPK promotes PD-L1 expression in RCC by enhancing TGF-β/Smad pathway activity The expression of different proteins in different cells was detected by immunoblot analysis. (A) Detection of TOPK binding to Smad4 by immunoprecipitation. (B-C) The expression of TOPK and Smad4 in TOPK knockdown 786-O cells(B) and overexpressed Caki-1 cells(C) were detected, respectively. (D) The expression of TOPK, p-TOPK(Thr9), Smad4, p-Smad4 (Thr276), Smad2/3, p-Smad2/3 (S465, 467/S423, 425), and PD-L1 was detected in 293T cells with or without TGF-β1 (5 ng/ml, 2h). (E-F) The expression of TOPK, p-TOPK(Thr9), Smad4, p-Smad4 (Thr276), Smad2/3, p-Smad2/3 (S465, 467/S423, 425), and PD-L1 was detected in TOPK knockdown (E) or overexpressed cells(F) with or without TGF-β1 (5 ng/ml, 2h). (G) Expression of TOPK, p-TOPK(Thr9), Smad4, p-Smad4 (Thr276), Smad2/3, p-Smad2/3 (S465, 467/S423, 425), and PD-L1 was detected in TOPK overexpressing cells alone or in combination with TGF-β1 (5 ng/mL, 2h) and OTS964 (10nM, 24h). See also Figure S4.

4). Does Chronic Pancreatitis in Growing Pigs Lead to Articular Cartilage Degradation and Alterations in Subchondral Bone?. International journal of molecular sciences (PubMed: 38396667) [IF=5.6]

5). MYOC Promotes the Differentiation of C2C12 Cells by Regulation of the TGF-β Signaling Pathways via CAV1. Geobiology (PubMed: 34356541) [IF=3.7]

Application: WB    Species: Mice    Sample: C2C12 cell

Figure 4 Effects of CAV1 on C2C12 cell differentiation. (A). Change in CAV1, MYH2, and MYOG expression levels at different stages of C2C12 cell differentiation. (B). Grayscale scan of CAV1 in Figure A. (C). Grayscale scan of MYH2 in Figure A. (D). Grayscale scan of MYOG in Figure A. (E). Immunofluorescence was used to measure the changes in CAV1 expression levels and the morphology of myotubes at various stages of C2C12 differentiation. CAV1 was labeled green using FITC and the nucleus blue using DAPI. (F). Laser confocal microscopy demonstrating the location of CAV1 expression, stained red using RBFITC, and nuclei blue using DAPI. (G). Changes in CAV1, MYH2, and MYOG expression levels after siRNA transfection for 48 h and 72 h. (H). Grayscale scan of CAV1 in Figure G. (I). Grayscale scan of MYH2 in Figure G. (J). Grayscale scan of MYOG in Figure G. (K). Changes in the fusion rate of C2C12 cells in mice after CAV1 inhibition by immunofluorescence detection. (L). Myotubule fusion rate in Figure K. (M). Change in CAV1, P-Smad2, and P-Smad4 expression after siRNA fragment S1 transfection of CAV1. (N). Grayscale scan of CAV1 in Figure M. (O). Grayscale scan of P-Smad2 in Figure M. (P). Grayscale scan of P-Smad4 in Figure M. (** p values < 0.01, * p values < 0.05) (n = 3).

6). Octanoyl esterification of low molecular weight sulfated galactan enhances the cellular uptake and collagen expression in fibroblast cells. Biomedical reports (PubMed: 37954636) [IF=2.3]

Application: WB    Species: Human    Sample: L929 fibroblasts

Figure 7 Effect of Oct-LMSG on Smad signaling protein expression in L929 fibroblasts. Smad2/3, Smad4, p-Smad2/3 and p-Smad4 protein levels were detected by western blot assay after treatment with either Oct-LMSG (100 µg/ml) or TGF-β (10 ng/ml) for 6, 12 and 24 h relative to β-actin. The values are expressed as the mean ± standard error of the mean of three independent experiments. *P

限制条款

产品的规格、报价、验证数据请以官网为准,官网链接:www.affbiotech.com | www.affbiotech.cn(简体中文)| www.affbiotech.jp(日本語)

产品的数据信息为Affinity所有,未经授权不得收集Affinity官网数据或资料用于商业用途,对抄袭产品数据的行为我们将保留诉诸法律的权利。

产品相关数据会因产品批次、产品检测情况随时调整,如您已订购该产品,请以订购时随货说明书为准,否则请以官网内容为准,官网内容有改动时恕不另行通知。

Affinity保证所销售产品均经过严格质量检测。如您购买的商品在规定时间内出现问题需要售后时,请您在Affinity官方渠道提交售后申请。

产品仅供科学研究使用。不用于诊断和治疗。 

产品未经授权不得转售。

Affinity Biosciences将不会对在使用我们的产品时可能发生的专利侵权或其他侵权行为负责。Affinity Biosciences, Affinity Biosciences标志和所有其他商标所有权归Affinity Biosciences LTD.