Overview

  • Product nameAnti-Smad2 antibodySee all Smad2 primary antibodies ...
  • Description
    Rabbit polyclonal to Smad2
  • Tested applicationsIHC-Fr, ICC/IF, WB, IHC-P more details
  • Species reactivity
    Reacts with: Mouse, Rat, Chicken, Cow, Dog, Human, Chimpanzee
    Predicted to work with: a wide range of other species
  • Immunogen

    Amino acids 234-249 (DQQLNQSMDTGSPAEL) of human SMAD2 protein.

  • Positive control
    • Rat or mouse muscle lysate This antibody gave a positive result when used in the following formaldehyde fixed cell lines: PC3.

Properties

Applications

Our Abpromise guarantee covers the use of ab47083 in the following tested applications.

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

Application Notes
IHC-Fr 1/30. PubMed: 19556713
ICC/IF Use a concentration of 5 µg/ml.
WB Use a concentration of 2 - 4 µg/ml. Detects a band of approximately 57 kDa (predicted molecular weight: 52 kDa).
IHC-P Use at an assay dependent concentration. PubMed: 20962090

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.
  • Target information above from: UniProt accession Q15796 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links
  • Alternative names
    • hMAD 2 antibody
    • hMAD-2 antibody
    • hSMAD2 antibody
    • JV18 1 antibody
    • JV18 antibody
    • JV18-1 antibody
    • JV181 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 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 images

  • ICC/IF image of ab47083 stained PC3 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab47083 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • All lanes : Anti-Smad2 antibody (ab47083) at 3 µg/ml

    Lane 1 : rat muscle lysate
    Lane 2 : rat muscle lysate in the presence of immunizing peptide.


    Predicted band size : 52 kDa
    Observed band size : 57 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 21 kDa,70 kDa. We are unsure as to the identity of these extra bands.

References for Anti-Smad2 antibody (ab47083)

This product has been referenced in:
  • Dupuis LE  et al. Insufficient versican cleavage and Smad2 phosphorylation results in bicuspid aortic and pulmonary valves. J Mol Cell Cardiol 60:50-9 (2013). IHC ; Mouse . Read more (PubMed: 23531444) »
  • Xu S  et al. Marek's disease virus type 1 microRNA miR-M3 suppresses cisplatin-induced apoptosis by targeting Smad2 of the transforming growth factor beta signal pathway. J Virol 85:276-85 (2011). WB, IHC-P ; Chicken . Read more (PubMed: 20962090) »

See all 3 Publications for this product

Product Wall

Application Western blot
Loading amount 40 µg
Gel Running Conditions Reduced Non-Denaturing (Native) (12)
Sample Mouse Tissue lysate - whole (Gastrocnemius (Muscle))
Specification Gastrocnemius (Muscle)
Blocking step Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Username

Mr. FRANCESCO ELIA MARINO

Verified customer

Submitted Mar 17 2014

Application Western blot
Sample Cow Cell lysate - whole cell (Disc cell)
Loading amount 30 µg
Specification Disc cell
Treatment TGFb1 recombinant protein 2ng/mL for 30min / TFb1 and ALK1 inhibitor concentration dependent for 30min
Gel Running Conditions Reduced Denaturing (12% gel)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jun 29 2012

Thank for your enquiry. We have not tested this antibody with recombinant SMAD proteins nor otherwise evaluated the cross-reactivity or lack thereof to other SMAD proteins. However, in doing a BLAST search using the immunogen sequence for this prod...

Read More

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