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RecombinantRabMAb

Recombinant Anti-Smad4 antibody [SP306] - C-terminal (ab217267)

Submit a review Submit a question References (1)
Flow Cytometry (Intracellular) - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)
  • Immunocytochemistry/ Immunofluorescence - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)
  • Flow Cytometry (Intracellular) - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)
  • Flow Cytometry (Intracellular) - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)
  • Flow Cytometry (Intracellular) - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)
  • Immunocytochemistry/ Immunofluorescence - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)
  • Immunocytochemistry/ Immunofluorescence - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)
  • Flow Cytometry (Intracellular) - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)
  • Anti-Smad4 antibody [SP306] - C-terminal (ab217267)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [SP306] to Smad4 - C-terminal
  • Suitable for: IHC-P, Flow Cyt (Intra), ICC/IF
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 Carrier Free

Overview

  • Product name

    Anti-Smad4 antibody [SP306] - C-terminal
    See all Smad4 primary antibodies

  • Description

    Rabbit monoclonal [SP306] to Smad4 - C-terminal

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, Flow Cyt (Intra), ICC/IFmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • Flow Cyt (intra): NIH/3T3, HepG2, HAP1 cells and C6 cells. ICC/IF: NIH/3T3, HepG2, and C6 cells. IHC-P: Human placenta, pancreas, pancreatic adenocarcinoma, kidney, renal cell carcinoma, colon and colon adenocarcinoma tissues.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    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 FOR RESEARCH USE ONLY. For commercial use, please contact partnerships@abcam.com.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab217267 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
IHC-P
1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt (Intra)
1/450.
ICC/IF
1/50.
Notes
IHC-P
1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt (Intra)
1/450.
ICC/IF
1/50.

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

    see all

  • 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

  • Flow Cytometry (Intracellular) - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)
    Flow Cytometry (Intracellular) - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)

    Flow cytometry overlay histogram showing wild-type Hap1 (green line) and SMAD4 knockout Hap1 stained with ab217267 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab217267) (1x 106 in 100μl at 0.008 μg/ml (1/265000)) for 30min at 22°C.

    The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C

    Isotype control antibody Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control was used at the same concentration and conditions as the primary antibody (wild-type Hap1 - black line, SMAD4 knockout Hap1 - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).

    Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

    This antibody gave a positive signal in Hap1 Fixed with 4% formaldehyde (10 min) / permeabilised with 0.1% PBS-Triton X-100 for 15 min under the same conditions.

     

  • Immunocytochemistry/ Immunofluorescence - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)
    Immunocytochemistry/ Immunofluorescence - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)
    Immunocytochemistry/ Immunofluorescence analysis of NIH/3T3 (mouse embryonic fibroblast) cells labeling Smad4 with purified ab217267 at 1:50 (9 µg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)

    Immunohistochemical analysis of formalin-fixed, paraffin-embedded human pancreas tissue labeling Smad4 with ab217267 at 1/100 dilution.

  • Flow Cytometry (Intracellular) - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)
    Flow Cytometry (Intracellular) - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)

    Intracellular Flow Cytometry analysis of C6 (Rat glial tumor glial cell) cells labeling Smad4 with purified ab217267 at 1/450 dilution (1.01 µg/ml) Red. Cells were fixed with 4% paraformaldehyde . A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) / Black. Unlabeled control - Unlabelled cells / Blue.

     

  • Flow Cytometry (Intracellular) - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)
    Flow Cytometry (Intracellular) - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)

    Intracellular Flow Cytometry analysis of NIH/3T3 (Mouse embryonic fibroblast) cells labeling Smad4 with purified ab217267 at 1/450 dilution (1.01 µg/ml) Red. Cells were fixed with 4% paraformaldehyde . A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) / Black. Unlabeled control - Unlabelled cells / Blue.

     

  • Flow Cytometry (Intracellular) - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)
    Flow Cytometry (Intracellular) - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)

    Intracellular Flow Cytometry analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling Smad4 with purified ab217267 at 1/450 dilution (1.01 µg/ml) Red. Cells were fixed with 4% paraformaldehyde . A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) / Black. Unlabeled control - Unlabelled cells / Blue.

     

  • Immunocytochemistry/ Immunofluorescence - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)
    Immunocytochemistry/ Immunofluorescence - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)
    Immunocytochemistry/ Immunofluorescence analysis of C6 (rat glial tumor glial cell) cells labeling Smad4 with purified ab217267 at 1:50 (9 µg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
  • Immunocytochemistry/ Immunofluorescence - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)
    Immunocytochemistry/ Immunofluorescence - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)
    Immunocytochemistry/ Immunofluorescence analysis of HepG2 (human hepatocellular carcinoma epithelial cell) cells labeling Smad4 with purified ab217267 at 1:50 (9 µg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
  • Flow Cytometry (Intracellular) - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)
    Flow Cytometry (Intracellular) - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)

    Intracellular flow cytometric analysis of NIH/3T3 (Mouse embryo fibroblast cell line) cellslabeling Smad4 with ab217267 at 1/400 dilution for 30 minutes at 4° C (green), compared to a negative control cell of rabbit IgG (blue). 

     

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)

    Immunohistochemical analysis of formalin-fixed, paraffin-embedded human placenta tissue labeling Smad4 with ab217267 at 1/100 dilution.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)

    Immunohistochemical analysis of formalin-fixed, paraffin-embedded human pancreatic adenocarcinoma tissue labeling Smad4 with ab217267 at 1/100 dilution.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)

    Immunohistochemical analysis of formalin-fixed, paraffin-embedded human kidney tissue labeling Smad4 with ab217267 at 1/100 dilution.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)

    Immunohistochemical analysis of formalin-fixed, paraffin-embedded human renal cell carcinoma tissue labeling Smad4 with ab217267 at 1/100 dilution.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)

    Immunohistochemical analysis of formalin-fixed, paraffin-embedded human colon tissue labeling Smad4 with ab217267 at 1/100 dilution.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [SP306] - C-terminal (ab217267)

    Immunohistochemical analysis of formalin-fixed, paraffin-embedded human colon adenocarcinoma tissue labeling Smad4 with ab217267 at 1/100 dilution.

  • Anti-Smad4 antibody [SP306] - C-terminal (ab217267)
    Anti-Smad4 antibody [SP306] - C-terminal (ab217267)

References (1)

Publishing research using ab217267? Please let us know so that we can cite the reference in this datasheet.

ab217267 has been referenced in 1 publication.

  • Kemper M  et al. Integrin alpha-V is an important driver in pancreatic adenocarcinoma progression. J Exp Clin Cancer Res 40:214 (2021). PubMed: 34174926

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