Validated using a knockout cell line
Recombinant
RabMAb

Anti-Smad4 antibody [EP618Y] - BSA and Azide free (ab215968)

Overview

  • Product name
    Anti-Smad4 antibody [EP618Y] - BSA and Azide free
    See all Smad4 primary antibodies
  • Description
    Rabbit monoclonal [EP618Y] to Smad4 - BSA and Azide free
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-Pmore details
    Unsuitable for: Flow Cyt,ICC/IF or IP
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Smad4 aa 500 to the C-terminus (C terminal).

  • Positive control
    • WB: NIH/3T3, PC-12, Ramos and SH-SY5Y cell lysates. IHC-P: human lung carcinoma and breast carcinoma tissues.
  • General notes

    ab215968 is a PBS-only buffer formulated version of ab40759, containing no BSA or sodium azide, ideal for antibody labeling. Please refer to ab40759 for information on protocols, dilutions, and image data.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab215968 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB Use at an assay dependent concentration. Detects a band of approximately 60 kDa (predicted molecular weight: 65 kDa).

 

IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

  • Application notes
    Is unsuitable for Flow Cyt,ICC/IF or IP.
  • Target

    • Function
      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 syngernistic transcriptional activity in response to TGF-beta. May act as a tumor suppressor.
    • Involvement in disease
      Defects 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 similarities
      Belongs to the dwarfin/SMAD family.
      Contains 1 MH1 (MAD homology 1) domain.
      Contains 1 MH2 (MAD homology 2) domain.
    • Domain
      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.
    • Post-translational
      modifications
      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. Deubiqitination by USP9X restores its competence to mediate TGF-beta signaling.
    • Cellular localization
      Cytoplasm. Nucleus. Cytoplasmic in the absence of ligand. Migrates to the nucleus when complexed with R-SMAD.
    • Information by UniProt
    • Database links
    • Alternative names
      • (Small) Mothers Against Decapentaplegic antibody
      • Deleted in Pancreatic Carcinoma 4 antibody
      • Deleted in Pancreatic Carcinoma antibody
      • Deleted in pancreatic carcinoma locus 4 antibody
      • Deletion target in pancreatic carcinoma 4 antibody
      • DPC 4 antibody
      • DPC4 antibody
      • hSMAD4 antibody
      • JIP antibody
      • MAD homolog 4 antibody
      • MAD mothers against decapentaplegic Drosophila homolog 4 antibody
      • MAD mothers against decapentaplegic homolog 4 antibody
      • MADH 4 antibody
      • MADH4 antibody
      • Med antibody
      • Medea antibody
      • Mothers against decapentaplegic homolog 4 antibody
      • Mothers against decapentaplegic, Drosophila, homolog of, 4 antibody
      • Mothers against DPP homolog 4 antibody
      • MYHRS antibody
      • OTTHUMP00000163548 antibody
      • SMA- and MAD-related protein 4 antibody
      • SMAD 4 antibody
      • SMAD family member 4 antibody
      • SMAD mothers against DPP homolog 4 antibody
      • SMAD4 antibody
      • SMAD4_HUMAN antibody
      see all

    Images

    • This IHC data was generated using the same anti-Smad4 antibody clone, EP618Y, in a different buffer formulation (cat# ab40759).

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labelling Smad4 with purified ab40759 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    • This WB data was generated using the same anti-Smad4 antibody clone, EP618Y, in a different buffer formulation (cat# ab40759).

      Lane 1: Wild type HAP1 whole cell lysate (20 µg)
      Lane 2: SMAD4 knockout  HAP1 whole cell lysate (20 µg)
      Lane 3: HepG2 whole cell lysate (20 µg)
      Lane 4: Jurkat whole cell lysate (20 µg)

      Lanes 1 - 4: Merged signal (red and green). Green - ab40759 observed at 60 kDa. Red - loading control, ab9484, observed at 37 kDa.

      ab40759 was shown to specifically react with SMAD4 in wild type HAP1 cells. No band was observed when SMAD4 knockout HAP1 samples were used. Wild-type and SMAD4 knockout samples were subjected to SDS-PAGE. Ab40759 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    References

    This product has been referenced in:
    • Shi S  et al. Metabolic tumor burden is associated with major oncogenomic alterations and serum tumor markers in patients with resected pancreatic cancer. Cancer Lett 360:227-33 (2015). IHC ; Human . Read more (PubMed: 25687883) »
    • Chen RY  et al. Effect of oridonin-mediated hallmark changes on inflammatory pathways in human pancreatic cancer (BxPC-3) cells. World J Gastroenterol 20:14895-903 (2014). WB ; Human . Read more (PubMed: 25356049) »

    See all 20 Publications for this product

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