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, Flow Cyt, ICC/IFmore 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 notes

    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.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

     

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 - 1/2000. Detects a band of approximately 58 kDa (predicted molecular weight: 58 kDa).

For unpurified, use 1/1000.

Flow Cyt 1/110.

For unpurified, use 1/70.

ICC/IF 1/300.
  • Application notesIs unsuitable for IHC-P or 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

    • Immunofluorescent staining of A673 cells, fixed with 4% PFA, using purified ab33875 at a dilution of 1/300. An Alexa Fluor® 555 goat anti-rabbit was used at 1/200. The negative control is shown in the bottom right hand panel.

    • Anti-Smad2 antibody [EP567Y] (ab33875) at 1/1000 dilution (Purified) + RAW264.7 at 10 µg

      Secondary
      HRP goat anti-rabbit (H+L) at 1/1000 dilution

      Predicted band size : 58 kDa

      5% NFDM/TBST dilution buffer

    • Anti-Smad2 antibody [EP567Y] (ab33875) at 1/2000 dilution (Purified) + Jurkat cell lysate at 10 µg

      Secondary
      HRP goat anti-rabbit (H+L) at 1/1000 dilution

      Predicted band size : 58 kDa
      Additional bands at : 58 kDa. We are unsure as to the identity of these extra bands.

      5% NFDM/TBST dilution buffer

    • Overlay histogram showing Jurkat cells stained with purified ab33875 (pink line) at a dilution of 1/110. The cells were fixed with 2% PFA. FITC goat anti-rabbit was used at a dilution of 1/150 and rabbit monoclonal IgG was used as the isotype control (green line).

    • Anti-Smad2 antibody [EP567Y] (ab33875) at 1/500 dilution + RAW264.7 cell lysate at 10 µg

      Secondary
      HRP goat anti-rabbit (H+L) at 1/1000 dilution

      Predicted band size : 58 kDa
      Additional bands at : 58 kDa. We are unsure as to the identity of these extra bands.

      5% NFDM/TBST dilution buffer

    • Anti-Smad2 antibody [EP567Y] (ab33875) at 1/1000 dilution (Unpurified) + Jurkat cell lysate at 10 µg

      Secondary
      HRP goat anti-rabbit (H+L) at 1/1000 dilution

      Predicted band size : 58 kDa
      Additional bands at : 58 kDa. We are unsure as to the identity of these extra bands.

      5% NFDM/TBST dilution buffer

    • All lanes : Anti-Smad2 antibody [EP567Y] (ab33875) at 1/3000 dilution (Unpurified)

      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 unpurified 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"