Rabbit Recombinant Monoclonal SMAD4 antibody - conjugated to HRP. Suitable for WB and reacts with Human samples.
pH: 7.4
Preservative: 0.1% Proclin 300 Solution
Constituents: PBS, 30% Glycerol (glycerin, glycerine), 1% BSA
IHC-P | WB | |
---|---|---|
Human | Not recommended | Tested |
Mouse | Not recommended | Predicted |
Rat | Not recommended | Predicted |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Species Mouse | Dilution info - | Notes - |
Species Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/5000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info - | 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 HRP. Suitable for WB and reacts with Human samples.
pH: 7.4
Preservative: 0.1% Proclin 300 Solution
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.
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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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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Terms & Conditions.
ab195554 was shown to recognize Smad4 in wild-type HAP1 cells as signal was lost at the expected MW in SMAD4 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and SMAD4 knockout samples were subjected to SDS-PAGE. ab195554 and Alexa Fluor® 680 Anti-GAPDH antibody [mAbcam 9484] - Loading Control ab184095 (Mouse monoclonal [mAbcam 9484] to GAPDH - Loading Control (Alexa Fluor® 680) loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/20000 dilution respectively. The loading control was imaged using the Licor Odyssey CLx prior to blots being developed with ECL technique.
All lanes: Western blot - HRP Anti-Smad4 antibody [EP618Y] (ab195554) at 1/5000 dilution
Lane 1: Wild-type HAP1 whole cell lysate at 20 µg
Lane 2: SMAD4 knockout HAP1 whole cell lysate at 20 µg
Predicted band size: 60 kDa
Observed band size: 65 kDa
Exposure time: 12min
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab195554 overnight at 4°C. Antibody binding was visualised using ECL development solution ECL Substrate Kit (High Sensitivity) ab133406.
All lanes: Western blot - HRP Anti-Smad4 antibody [EP618Y] (ab195554) at 1/5000 dilution
All lanes: SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate at 10 µg
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 60 kDa, 82 kDa
Observed band size: 30 kDa, 47 kDa, 52 kDa, 54 kDa, 65 kDa, 82 kDa
Exposure time: 20min
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