Rabbit Recombinant Monoclonal SMAD4 antibody - conjugated to Alexa Fluor® 488. C-terminal. Suitable for IHC-P, ICC/IF and reacts with Human, Mouse samples.
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: PBS, 30% Glycerol (glycerin, glycerine), 1% BSA
IHC-P | ICC/IF | |
---|---|---|
Human | Tested | Expected |
Mouse | Expected | Tested |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/100 | Notes Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
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Species Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/100 | Notes This product gave a positive signal in NIH3T3 fixed with 4% formaldehyde (10 min). |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info Use at an assay dependent concentration. | Notes - |
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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 (PubMed:25514493). 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.
DPC4, MADH4, SMAD4, Mothers against decapentaplegic homolog 4, MAD homolog 4, Mothers against DPP homolog 4, Deletion target in pancreatic carcinoma 4, SMAD family member 4, SMAD 4, Smad4, hSMAD4
Rabbit Recombinant Monoclonal SMAD4 antibody - conjugated to Alexa Fluor® 488. C-terminal. Suitable for IHC-P, ICC/IF and reacts with Human, Mouse samples.
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: PBS, 30% Glycerol (glycerin, glycerine), 1% BSA
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company. The Alexa Fluor® dye included in this product is provided under an intellectual property license from Life Technologies Corporation. As this product contains the Alexa Fluor® dye, the purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). As this product contains the Alexa Fluor® dye the sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are sold for use in research. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@thermofisher.com.
Smad4 also known as DPC4 or MADH4 is a central protein in the TGF-beta signaling pathway with a molecular mass of approximately 60 kDa. It acts as a signal transducer that facilitates communication from the cell surface to the nucleus. Smad4 is broadly expressed in various tissues playing an important role in the regulation of cellular processes. It forms a complex with receptor-regulated Smads (R-Smads) to translocate to the nucleus where it influences gene transcription.
Smad4 influences cell proliferation differentiation and apoptosis by mediating signals from TGF-beta cytokines. It is part of the Smad protein family acting as a transcriptional controller. Upon TGF-beta receptor activation Smad4 forms complexes with Smad2 and Smad3 translocating to the nucleus to regulate genes imperative for cellular homeostasis. Its role in cell cycle regulation underlines its contribution to normal cellular functions and its potential involvement in disorders.
Smad4 operates within the TGF-beta pathway linking extracellular signals to nuclear transcription alterations. It participates in the regulation of epithelial-mesenchymal transition (EMT) a process important for development and tumor progression. In these pathways Smad4 interacts closely with Smad2 and Smad3 orchestrating various cellular responses to external stimuli through transcriptional management.
Smad4 is highly related to cancer and juvenile polyposis syndrome. Mutations or deletions in Smad4 disrupt its function contributing to the progression of pancreatic cancer and colorectal cancer among others. Within these contexts Smad4 connects strongly to other proteins like p21 and cyclin-dependent kinase inhibitors which are important in cell cycle arrest and impede tumor growth.
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We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
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Immunofluorescence staining of Smad4 - C-terminal staining in a section of formalin-fixed paraffin-embedded normal human colon*.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6) in a Biocare Medical NxGen pressure cooker using retrieval settings of 110°C for 20 minutes. Non-specific protein-protein interactions were then blocked in TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 1% (w/v) BSA for 1h at room temperature. The section was then incubated overnight at +4°C in TBS containing 0.025% (v/v) Triton X-100 and 1% (w/v) BSA with ab267520 at 1/100 dilution (shown in green) and counterstained using Alexa Fluor® 647 Anti-Tubulin antibody [YOL1/34] - Microtubule Marker ab195884, Rat monoclonal to Tubulin (Alexa Fluor® 647), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Fluoromount®.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.
Immunofluorescence staining of Smad4 in NIH3T3 cells using ab267520. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab267520 at 1/100 dilution (shown in green) and Alexa Fluor® 647 Anti-Tubulin antibody [YOL1/34] - Microtubule Marker ab195884, Rat monoclonal to Tubulin (Alexa Fluor® 647), at 1/250 dilution (shown in ). Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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