Overview

  • Product nameAnti-Smad2 antibody [EP567Y]See all Smad2 primary antibodies ...
  • Description
    Rabbit monoclonal [EP567Y] to Smad2
  • SpecificityThis antibody detects a region about 40AA before the MH2 region(not the MH2 region itself)
  • Tested applicationsWB, IHC-P, ICC, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    A synthetic peptide corresponding to residues surrounding the MH2 domain of human Smad2.

  • Positive control
    • Jurkat cell lysate and human adenocarcinoma of uterus
  • General notesProduced under U.S. Patent No. 5,675,063.

Properties

Applications

Our Abpromise guarantee covers the use of ab33875 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 1/1000. Detects a band of approximately 58 kDa (predicted molecular weight: 58 kDa).
IHC-P Use at an assay dependent dilution.
ICC 1/250 - 1/500.
Flow Cyt 1/20 - 1/100.
  • Application notesIs unsuitable for IP.
  • Target

    • FunctionReceptor-regulated SMAD (R-SMAD) that is an intracellular signal transducer and transcriptional modulator activated by TGF-beta (transforming growth factor) and activin type 1 receptor kinases. Binds the TRE element in the promoter region of many genes that are regulated by TGF-beta and, on formation of the SMAD2/SMAD4 complex, activates transcription. May act as a tumor suppressor in colorectal carcinoma.
    • Tissue specificityExpressed at high levels in skeletal muscle, heart and placenta.
    • Sequence similaritiesBelongs to the dwarfin/SMAD family.
      Contains 1 MH1 (MAD homology 1) domain.
      Contains 1 MH2 (MAD homology 2) domain.
    • Post-translational
      modifications
      Phosphorylated on one or several of Thr-220, Ser-245, Ser-250, and Ser-255. In response to TGF-beta, phosphorylated on Ser-465/467 by TGF-beta and activin type 1 receptor kinases. Able to interact with SMURF2 when phosphorylated on Ser-465/467, recruiting other proteins, such as SNON, for degradation. In response to decorin, the naturally occurring inhibitor of TGF-beta signaling, phosphorylated on Ser-240 by CaMK2. Phosphorylated by MAPK3 upon EGF stimulation; which increases transcriptional activity and stability, and is blocked by calmodulin.
      In response to TGF-beta, ubiquitinated by NEDD4L; which promotes its degradation.
      Acetylated on Lys-19 by coactivators in response to TGF-beta signaling, which increases transcriptional activity. Isoform short: Acetylation increases DNA binding activity in vitro and enhances its association with target promoters in vivo. Acetylation in the nucleus by EP300 is enhanced by TGF-beta.
    • Cellular localizationCytoplasm. Nucleus. Cytoplasmic and nuclear in the absence of TGF-beta. On TGF-beta stimulation, migrates to the nucleus when complexed with SMAD4. On dephosphorylation by phosphatase PPM1A, released from the SMAD2/SMAD4 complex, and exported out of the nucleus by interaction with RANBP1.
    • Information by UniProt
    • Database links
    • Alternative names
      • hMAD 2 antibody
      • hMAD-2 antibody
      • hSMAD2 antibody
      • JV18 1 antibody
      • JV18 antibody
      • JV18 antibody
      • JV18-1 antibody
      • JV181 antibody
      • MAD antibody
      • MAD antibody
      • MAD homolog 2 antibody
      • MAD Related Protein 2 antibody
      • Mad-related protein 2 antibody
      • MADH2 antibody
      • MADR2 antibody
      • MGC22139 antibody
      • MGC34440 antibody
      • Mother against DPP homolog 2 antibody
      • Mothers Against Decapentaplegic Homolog 2 antibody
      • Mothers Against Decapentaplegic Homolog 2 antibody
      • mothers against DPP homolog 2 antibody
      • OTTHUMP00000163489 antibody
      • Sma and Mad related protein 2 antibody
      • SMAD 2 antibody
      • SMAD antibody
      • SMAD family member 2 antibody
      • SMAD, mothers against DPP homolog 2 antibody
      • SMAD2 antibody
      • SMAD2_HUMAN antibody
      see all

    Anti-Smad2 antibody [EP567Y] images

    • Anti-Smad2 antibody [EP567Y] (ab33875) at 1/1000 dilution + Jurkat cell lysate

      Predicted band size : 58 kDa
      Observed band size : 58 kDa
    • ab33875, at a 1/50 dilution,staining human adenocarcinoma of uterus by Immunohistochemistry, Paraffin embedded tissue.
    • All lanes : Anti-Smad2 antibody [EP567Y] (ab33875) at 1/3000 dilution

      Lane 1 : HGL-5 cells - no additives
      Lane 2 : Cells + activin A
      Lane 3 : Cells + cyclophilin siRNA Control
      Lane 4 : Cells + activin A + cyclophilin
      Lane 5 : non-targeting, negative control for siRNA
      Lane 6 : Cells + activin A, negative control for siRNA
      Lane 7 : Cells + siRNA Smartpool
      Lane 8 : Cells + activin A + siRNA Smartpool
      Lane 9 : Jurkat cells

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/50000 dilution

      Performed under reducing conditions.

      Predicted band size : 58 kDa
      Observed band size : 58 kDa

      This image is courtesy of an Abreview submitted by Mrs Birte Jablonski

      See Abreview

    • Overlay histogram showing PC3 cells stained with ab33875 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab33875, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in PC3 cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

    References for Anti-Smad2 antibody [EP567Y] (ab33875)

    This product has been referenced in:
    • Gallo EM  et al. Angiotensin II-dependent TGF-ß signaling contributes to Loeys-Dietz syndrome vascular pathogenesis. J Clin Invest 124:448-60 (2014). IF . Read more (PubMed: 24355923) »
    • Zieba A  et al. Intercellular variation in signaling through the TGF-ß pathway and its relation to cell density and cell cycle phase. Mol Cell Proteomics 11:M111.013482 (2012). Read more (PubMed: 22442258) »

    See all 5 Publications for this product

    Product Wall

    Thank you for contacting us.

    We do have a number of anti-SMAD2 and anti-SMAD3 antibodies which we guarantee in IHC-P on mouse. We to don't have the opportunity to test specific products against one another in each application but based on ...

    Read More
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Sample Mouse Cell lysate - whole cell (c3h10t1/2)
    Loading amount 25 µg
    Specification c3h10t1/2
    Treatment TGFbeta 15 and 30 minutes
    Gel Running Conditions Reduced Denaturing (4-12% gradient)
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Dr. Serban Georgescu

    Verified customer

    Submitted Mar 03 2008

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Sample Human Cell lysate - whole cell (Human cell line HGL-5)
    Loading amount 15 µg
    Specification Human cell line HGL-5
    Blocking step Milk as blocking agent for 15 hour(s) and 0 minute(s) · Concentration: 5%
    Username

    Mrs. Birte Münchow

    Verified customer

    Submitted Oct 17 2006

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"