Smad4 Antibody (YA4241)(PBS only)
(Synonyms: JIP; DPC4; MADH4; SMAD4)Smad4 Antibody (YA4241) is a Mouse-derived and non-conjugated IgG1 monoclonal antibody, targeting to Smad4.
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Host:
Mouse
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Isotype:
IgG
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Application:
WB, IHC-P, ICC/IF, FC, ELISA
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Reactivity :
Human, Mouse
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Formulation:
Supplied in PBS, pH 7.4.
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Conjugation:
Non-conjugated
To place orders, for customer services and technical support, please contact: MedChemExpress USA
Tel: 609-228-6898 E-mail: [email protected] [email protected]
Applications
| Application |
WB
WB: Western Blot
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IHC-P
IHC-P: Immunohistochemistry-Paraffin
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ICC/IF
ICC/IF: Immunocytochemistry/
Immunofluorescence |
FC
FC: Flow Cytometry
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ELISA
ELISA: Enzyme Linked Immunosorbent Assay
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|---|---|---|---|---|---|
| Dilution Ratio | 1:500-1:2000 | 1:200-1:1000 | 1:200-1:1000 | 1:200-1:400 | 1:10000 |
Product Details
Smad4 Antibody (YA4241) is a Mouse-derived and non-conjugated IgG1 monoclonal antibody, targeting to Smad4.
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Host Mouse
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Species ReactivityHuman, Mouse
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Observed Molecular WeightObserved band size: 65 kDa
Note: Due to possible protein modifications or aggregation, the molecular weight should be confirmed by actual measurement, and the predicted value is for reference only.
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Calculated Molecular Weight Predicted band size: 60 kDa
Purified recombinant fragment of human SMAD4 aa 336-552.
affinity purified.
Non-conjugated
IgG
Product Properties
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Appearance
Solution
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Formulation
Supplied in PBS, pH 7.4.
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Storage & Stability
Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.
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Shipping
Shipping with blue ice.
Verification Images
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Western blot analysis of extracts from A431(lane 2(20μg), K562 (lane 3(20μg) and HepG2 (lane 4(20μg) using SMAD4 (30091) Mouse mAb. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST for 2 hour at room temperature. The primary antibody (1/1000) and Loading control antibody (Beta Actin, HY-P80438, 1/10000) was used in 5% non-fat milk in TBST at 4°C overnight. Goat Anti-Mouse IgG-HRP Secondary Antibody (1/10000) was used for 1 hour at room temperature. -
Immunohistochemical analysis of paraffin-embedded Mouse lung tissue using SMAD4 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (30091, 1/500) in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded Mouse lung tissue using SMAD4 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (30091, 1/500) in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunocytochemistry analysis of NIH/3T3 cells labeling SMAD4 with SMAD4 Antibody (HY-P84544)at 1/200 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with SMAD4 Antibody (HY-P84544) at 1/200 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Mouse IgG H&L（HY-P8005, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue). -
Immunocytochemistry analysis of NIH/3T3 cells labeling SMAD4 with SMAD4 Antibody (HY-P84544) at 1/500 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with SMAD4 Antibody (HY-P84544) at 1/500 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Mouse IgG H&L（HY-P8005, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
Background
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Function
SMAD4 belongs to the dwarfin/SMAD family. SMAD is a group of proteins that carry extracellular signals directly to the nucleus. Mammalian SMAD can be divided into three categories: receptor-regulated SMAD (R-SMAD), common-mediated SMAD (Co-SMAD), and inhibitory SMAD (I-SMAD) . (1) R-SMAD included SMAD1, SMAD2, SMAD3, SMAD5 and SMAD8/9, among which Smad2/3 mediated signal transduction of TGF-β subfamily members. SMAD1/5/8 mediates signaling from members of the bone morphogenetic proteins (BMP) subfamily. (2) Co-SMAD includes SMAD4, which is the central mediator of TGF-β and BMP signaling pathways. (3) I-SMAD, including SMAD6 and SMAD7, inhibits receptor-mediated phosphorylation of R-SMAD, thereby preventing the formation of complexes with Co SMAD. SMAD4 plays an important role in cellular signaling of bone development (from chondrocyte precursors to mature osteocytes) and tissue homeostasis. Specifically, TGF-β signals upstream to activate SMAD4 and R-SMAD (i.e. SMAD2/3) to interact and form oligomer complexes that regulate the expression of target genes. However, SMAD3/4 mainly induced chondrogenesis of human bone marrow mesenchymal stem cells. Abnormal expression of SMAD4 can inhibit cartilage production in animal models, and even lead to uncontrolled cell growth and tumor induction. SMAD4 also specifically targets stem cells and may be involved in self-renewal of hematopoietic stem cells. At the same time, SMAD4 regulates the cell polarity of chondrocytes, affecting the shape, size, migration and orientation of chondrocytes. It also promotes cell migration, adhesion and cytoskeletal tissue generation in different cells through the SMAD4/TGF-β pathway. The SMAD4/TGF-β pathway is also extensively regulated by classical pathways such as MAPK, PI3K/AKT, and WNT/β-catenin to form complex networks. SMAD4 also regulates apoptosis, leading to follicular atresia by inhibiting granulosa cell apoptosis. In colorectal cancer cells, TGF-β induces Smad4-dependent epithelial-mesenchymal transformation, which leads to apoptosis. The protective effect of SMAD4 on apoptosis is also the key to the differentiation and proliferation of chondrocytes. In addition, apoptosis is crucial for differentiation and bone homeostasis, that is, osteoblast apoptosis promotes osteoclast generation and bone resorption, which is an important process of bone homeostasis. SMAD4 also controls gene regulatory networks in early limb buds, and chondrocyte specific Smad4-KO mice exhibit dwarfism and impaired growth plate tissue. SMAD4 protein also inhibited epithelial cell proliferation and decreased angiogenesis and increased vascular permeability, showing antitumor effects[1][2].
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Subcellular Localization
Cytoplasm; Nucleus
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Subunit
Monomer; in the absence of TGF-beta activation (PubMed:9670020). Heterotrimer; on TGF-beta activation (PubMed:15799969).
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SwissProt ID
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Synonyms
JIP; DPC4; MADH4; SMAD4
Documentation
References
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