Key features and details
- Rabbit polyclonal to Smad4
- Suitable for: IP, IHC-P, WB
- Reacts with: Mouse, Human
- Isotype: IgG
Product nameAnti-Smad4 antibody
See all Smad4 primary antibodies
DescriptionRabbit polyclonal to Smad4
Tested applicationsSuitable for: IP, IHC-P, WBmore details
Species reactivityReacts with: Mouse, Human
Predicted to work with: Rat, Cow, Pig
- WB: SH-SY5Y, 3T3 and HepG2 whole cell lysate. IHC-P: Human colon cancer and kidney tissue. IP: Jurkat cells.
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In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
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Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage bufferpH: 7.40
Constituents: PBS, 50% Glycerol, 0.03% Proclin 300
Concentration information loading...
PurityProtein G purified
Purification notesProtein >95%.
Our Abpromise guarantee covers the use of ab236321 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IP||1/200 - 1/2000.|
|IHC-P||1/20 - 1/200.|
|WB||1/500 - 1/2000. Predicted molecular weight: 60 kDa.|
FunctionCommon 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 syngernistic transcriptional activity in response to TGF-beta. May act as a tumor suppressor.
Involvement in diseaseDefects in SMAD4 are a cause of pancreatic cancer (PNCA) [MIM:260350].
Defects in SMAD4 are a cause of juvenile polyposis syndrome (JPS) [MIM:174900]; also known as juvenile intestinal polyposis (JIP). JPS is an autosomal dominant gastrointestinal hamartomatous polyposis syndrome in which patients are at risk for developing gastrointestinal cancers. The lesions are typified by a smooth histological appearance, predominant stroma, cystic spaces and lack of a smooth muscle core. Multiple juvenile polyps usually occur in a number of Mendelian disorders. Sometimes, these polyps occur without associated features as in JPS; here, polyps tend to occur in the large bowel and are associated with an increased risk of colon and other gastrointestinal cancers.
Defects in SMAD4 are a cause of juvenile polyposis/hereditary hemorrhagic telangiectasia syndrome (JP/HHT) [MIM:175050]. JP/HHT syndrome phenotype consists of the coexistence of juvenile polyposis (JIP) and hereditary hemorrhagic telangiectasia (HHT) [MIM:187300] in a single individual. JIP and HHT are autosomal dominant disorders with distinct and non-overlapping clinical features. The former, an inherited gastrointestinal malignancy predisposition, is caused by mutations in SMAD4 or BMPR1A, and the latter is a vascular malformation disorder caused by mutations in ENG or ACVRL1. All four genes encode proteins involved in the transforming-growth-factor-signaling pathway. Although there are reports of patients and families with phenotypes of both disorders combined, the genetic etiology of this association is unknown.
Defects in SMAD4 may be a cause of colorectal cancer (CRC) [MIM:114500].
Sequence similaritiesBelongs to the dwarfin/SMAD family.
Contains 1 MH1 (MAD homology 1) domain.
Contains 1 MH2 (MAD homology 2) domain.
DomainThe 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.
modificationsMonoubiquitinated 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. Deubiqitination by USP9X restores its competence to mediate TGF-beta signaling.
Cellular localizationCytoplasm. Nucleus. Cytoplasmic in the absence of ligand. Migrates to the nucleus when complexed with R-SMAD.
- Information by UniProt
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All lanes : Anti-Smad4 antibody (ab236321) at 1/500 dilution
Lane 1 : SH-SY5Y (human neuroblastoma cell line from bone marrow) whole cell lysate
Lane 2 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate
All lanes : Goat polyclonal to rabbit IgG at 1/10000 dilution
Predicted band size: 60 kDa
Paraffin-embedded human kidney tissue stained for Smad4 using ab236321 at 1/100 dilution in immunohistochemical analysis.
Paraffin-embedded human colon cancer tissue stained for Smad4 using ab236321 at 1/100 dilution in immunohistochemical analysis.
Anti-Smad4 antibody (ab236321) at 1/500 dilution + HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate
Goat polyclonal to rabbit IgG at 1/10000 dilution
Predicted band size: 60 kDa
Smad4 was immunoprecipitated from 0.5 mg Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate with ab236321.
Lane 1: Rabbit control IgG instead of ab236321 in Jurkat whole cell lysate.
Lane 2: ab236321 IP in Jurkat whole cell lysate.
Lane 3: Jurkat whole cell lysate 10 µg (Input).
For western blotting, a HRP-conjugated Protein G antibody was used as the secondary antibody at 1/2000 dilution.
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab236321 has been referenced in 2 publications.
- Bonan NF et al. Inhibition of HGF/MET signaling decreases overall tumor burden and blocks malignant conversion in Tpl2-related skin cancer. Oncogenesis 8:1 (2019). PubMed: 30631034
- Stojnev S et al. Prognostic Impact of Canonical TGF-ß Signaling in Urothelial Bladder Cancer. Medicina (Kaunas) 55:N/A (2019). PubMed: 31238579