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RabMAb

Anti-Smad4抗体[EP618Y] (ab40759)

概述

  • 产品名称
    Anti-Smad4抗体[EP618Y]
    参阅全部 Smad4 一抗
  • 描述
    兔单克隆抗体[EP618Y] to Smad4
  • 经测试应用
    适用于: WB, IHC-Pmore details
    不适用于: Flow Cyt,ICC/IF or IP
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

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

  • 阳性对照
    • WB: NIH/3T3, PC-12, Ramos and SH-SY5Y cell lysates. IHC-P: human lung carcinoma and breast carcinoma tissues.
  • 常规说明

    This product is a recombinant rabbit monoclonal antibody.

     

    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.

     

    Alternative versions available:

    Anti-Smad4 antibody (HRP) [EP618Y] (ab195554)

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

性能

应用

Our Abpromise guarantee covers the use of ab40759 in the following tested applications.

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

应用 Ab评论 说明
WB 1/5000. Detects a band of approximately 60 kDa (predicted molecular weight: 65 kDa).Can be blocked with Human Smad4 peptide (ab228416).

For unpurified use at 1/1000 - 1/5000.

IHC-P 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

For unpurified use at 1/100 - 1/200.

  • 应用说明
    Is unsuitable for Flow Cyt,ICC/IF or IP.
  • 靶标

    • 功能
      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.
    • 疾病相关
      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].
    • 序列相似性
      Belongs to the dwarfin/SMAD family.
      Contains 1 MH1 (MAD homology 1) domain.
      Contains 1 MH2 (MAD homology 2) domain.
    • 结构域
      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.
    • 翻译后修饰
      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.
    • 细胞定位
      Cytoplasm. Nucleus. Cytoplasmic in the absence of ligand. Migrates to the nucleus when complexed with R-SMAD.
    • Information by UniProt
    • 数据库链接
    • 别名
      • (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

    图片



    • Predicted band size : 65 kDa

      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.

       

    • All lanes : Anti-Smad4 antibody [EP618Y] (ab40759) at 1/5000 dilution

      Lane 1 : NIH/3T3 cell lysate
      Lane 2 : Mouse embryo tissue lysate
      Lane 3 : Mouse skin tissue lysate
      Lane 4 : Mouse lung tissue lysate
      Lane 5 : PC-12 cell lysate
      Lane 6 : C6 cell lysate
      Lane 7 : Rat skin tissue lysate
      Lane 8 : Rat lung tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size : 65 kDa
      Observed band size : 60 kDa (why is the actual band size different from the predicted?)


      Exposure time : 3 minutes

      Blocking and dilution buffer: 5% NFDM/TBST.

    • All lanes : Anti-Smad4 antibody [EP618Y] (ab40759) at 1/5000 dilution

      Lane 1 : SW480 cell lysate
      Lane 2 : HepG2 cell lysate
      Lane 3 : Jurkat cell lysate
      Lane 4 : Human skin tissue lysate
      Lane 5 : Human lung tissue lysate
      Lane 6 : Human artery tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size : 65 kDa
      Observed band size : 60 kDa (why is the actual band size different from the predicted?)


      Exposure time : 30 seconds

      Blocking and dilution buffer: 5% NFDM/TBST.

    • All lanes : Anti-Smad4 antibody [EP618Y] (ab40759) at 1/5000 dilution (purified)

      Lane 1 : SH-SY5Y cell lysate
      Lane 2 : Ramos cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

      Predicted band size : 65 kDa
      Observed band size : 60 kDa (why is the actual band size different from the predicted?)

      Blocking and diluting buffer: 5% NFDM/TBST.

    • Anti-Smad4 antibody [EP618Y] (ab40759) at 1/10000 dilution (purified) + NIH/3T3 cell lysate at 20 µg

      Secondary
      Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

      Predicted band size : 65 kDa
      Observed band size : 60 kDa (why is the actual band size different from the predicted?)

      Blocking and diluting buffer: 5% NFDM/TBST.

    • Anti-Smad4 antibody [EP618Y] (ab40759) at 1/5000 dilution (purified) + PC-12 cell lysate at 10 µg

      Secondary
      Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

      Predicted band size : 65 kDa
      Observed band size : 60 kDa (why is the actual band size different from the predicted?)

      Blocking and diluting buffer: 5% NFDM/TBST.

    • Anti-Smad4 antibody [EP618Y] (ab40759) at 1/5000 dilution (unpurified) + SHSY5Y cell lysate at 10 µg

      Predicted band size : 65 kDa
    • 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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue labelling Smad4 with unpurified ab40759 at a 1/100 dilution.

    • Unpurified ab40759 staining Smad4 in rat femur tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% BSA for 20 minutes at 22°C; antigen retrieval was by heat mediation in a citrate buffer pH6.0. Samples were incubated with primary antibody (1/200 in blocking buffer) for 2 hours at 20°C. An undiluted HRP-conjugated goat anti-rabbit IgG polyclonal (1/250) was used as the secondary antibody.

      See Abreview

    文献

    This product has been referenced in:
    • Kohutek ZA  et al. An unusual genomic variant of pancreatic ductal adenocarcinoma with an indolent clinical course. Cold Spring Harb Mol Case Stud 3:N/A (2017). IHC-P ; Human . Read more (PubMed: 28679692) »
    • Zhou J  et al. MicroRNA-144 is regulated by CP2 and decreases COX-2 expression and PGE2 production in mouse ovarian granulosa cells. Cell Death Dis 8:e2597 (2017). Read more (PubMed: 28182010) »

    See all 35 Publications for this product

    客户评价及客户问答

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Liver)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Dako
    Permeabilization
    No
    Specification
    Liver
    Fixative
    Formaldehyde
    Username

    Abcam user community

    Verified customer

    提交于 Jun 01 2016

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (postnatal day 21 mouse ovary)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: 0.01M citric acide/sodium citrate
    Permeabilization
    No
    Specification
    postnatal day 21 mouse ovary
    Blocking step
    BSA as blocking agent for 45 minute(s) · Concentration: 5% · Temperature: 25°C
    Fixative
    4% PFA
    Username

    Abcam user community

    Verified customer

    提交于 Jan 05 2015

    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (A549)
    Permeabilization
    Yes - 0.25% Triton X-100/PBS
    Specification
    A549
    Fixative
    Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    提交于 Apr 11 2013

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Lung tissue)
    Specification
    Lung tissue
    Fixative
    HOPE
    Antigen retrieval step
    None
    Permeabilization
    No
    Username

    Abcam user community

    Verified customer

    提交于 Mar 27 2013

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Rat Tissue sections (femur)
    Specification
    femur
    Fixative
    Formaldehyde
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrated buffer, pH6.0
    Permeabilization
    No
    Blocking step
    BSA as blocking agent for 20 minute(s) · Concentration: 1% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    提交于 Aug 23 2012

    Thank you for your reply. I will be happy to mail the datasheets to you. Are you having trouble accessing the datasheets online? I only ask so that I can inform our IT department if there are problems. Could you also please send me the address that you...

    Read More

    The datasheets are accessible from the product pages on the abcam.com website. You could try and print them from there. If you still have difficulty, please let me know and I will mail the datasheets to you.

    Thank you for contacting Abcam. Please find attached the missing datasheets. If there is anything else I can help you with, please let me kn

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (lymphoblats)
    Loading amount
    40 µg
    Specification
    lymphoblats
    Gel Running Conditions
    Reduced Denaturing (4-12% Bis-Tris Gel, MOPS Buffer)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Username

    Dr. Lukas Rambousek

    Verified customer

    提交于 Dec 15 2010

    Application
    ChIP
    Sample
    Human Cell lysate - whole cell (MDA-MB-231 Cells treated with 2 ng/µl TGF-beta1)
    Specification
    MDA-MB-231 Cells treated with 2 ng/µl TGF-beta1
    Type
    Cross-linking (X-ChIP)
    Duration of cross-linking step: 0 second(s)
    Specification of the cross-linking agent: 1% Formaldehyde
    Detection step
    Real-time PCR
    Negative control
    Polyclonal anti-HA antibody (for detection of non-specific binding). The graph is shown as a "fold enrichment" relative to the average detected HA binding.
    Username

    Abcam user community

    Verified customer

    提交于 Feb 14 2007

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