Smad4 Antibody - #AF5247

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Product: Smad4 Antibody
Catalog: AF5247
Description: Rabbit polyclonal antibody to Smad4
Application: WB IHC IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Bovine, Horse, Sheep, Rabbit, Dog
Mol.Wt.: 65 kDa; 60kD(Calculated).
Uniprot: Q13485
RRID: AB_2837733

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 100ul $280 In stock
 200ul $350 In stock

Lead Time: Same day delivery

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Related Downloads
MS Report
HPLC Report
MSDS
Data Sheet
COA
Protocols
WB Handbook
IHC Protocol
IF/ICC Protocol

Product Info

Source:
Rabbit
Application:
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: 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.

Reactivity:
Human,Mouse,Rat
Prediction:
Pig(100%), Bovine(100%), Horse(100%), Sheep(100%), Rabbit(100%), Dog(100%)
Clonality:
Polyclonal
Specificity:
Smad4 Antibody detects endogenous levels of total Smad4.
RRID:
AB_2837733
Cite Format: Affinity Biosciences Cat# AF5247, RRID:AB_2837733.
Conjugate:
Unconjugated.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
Storage:
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.
Alias:

Fold/Unfold

(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;

Immunogens

Immunogen:
Uniprot:

Q13485(SMAD4_HUMAN) >>Visit HPA database.

Gene(ID):
SMAD4(4089)
Description:
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.
Sequence:
MDNMSITNTPTSNDACLSIVHSLMCHRQGGESETFAKRAIESLVKKLKEKKDELDSLITAITTNGAHPSKCVTIQRTLDGRLQVAGRKGFPHVIYARLWRWPDLHKNELKHVKYCQYAFDLKCDSVCVNPYHYERVVSPGIDLSGLTLQSNAPSSMMVKDEYVHDFEGQPSLSTEGHSIQTIQHPPSNRASTETYSTPALLAPSESNATSTANFPNIPVASTSQPASILGGSHSEGLLQIASGPQPGQQQNGFTGQPATYHHNSTTTWTGSRTAPYTPNLPHHQNGHLQHHPPMPPHPGHYWPVHNELAFQPPISNHPAPEYWCSIAYFEMDVQVGETFKVPSSCPIVTVDGYVDPSGGDRFCLGQLSNVHRTEAIERARLHIGKGVQLECKGEGDVWVRCLSDHAVFVQSYYLDREAGRAPGDAVHKIYPSAYIKVFDLRQCHRQMQQQAATAQAAAAAQAAAVAGNIPGPGSVGGIAPAISLSAAAGIGVDDLRRLCILRMSFVKGWGPDYPRQSIKETPCWIEIHLHRALQLLDEVLHTMPIADPQPLD

Predictions

Predictions:

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
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

PTMs - Q13485 As Substrate

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

Research Backgrounds

Function:

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.

PTMs:

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.

Subcellular Location:

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
Subunit Structure:

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.

Family&Domains:

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.

Research Fields

· 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)

References

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]
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). 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).
6). 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
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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