Overview

  • Product name
    Anti-Smad2 antibody [7A5]
    See all Smad2 primary antibodies
  • Description
    Mouse monoclonal [7A5] to Smad2
  • Tested applications
    Suitable for: ChIP, WB, IP, ELISA, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant full length human Smad2 protein purified from E.coli with His tag.

  • Positive control
    • HeLa, A431, NIH3T3 and PC12 cell lysates.
  • General notes

    This product was changed from ascites to tissue culture supernatant on 18th September 2017. Lot numbers higher than GR183929 will be from tissue culture supernatant. Please note that the dilutions may need to be adjusted accordingly.

Properties

Applications

Our Abpromise guarantee covers the use of ab71109 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
ChIP Use at an assay dependent concentration.
WB Use a concentration of 0.1 µg/ml. Detects a band of approximately 58 kDa (predicted molecular weight: 58 kDa).
IP Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab170192-Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

Target

  • Function
    Receptor-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 specificity
    Expressed at high levels in skeletal muscle, heart and placenta.
  • Sequence similarities
    Belongs 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 localization
    Cytoplasm. 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
    • Drosophila, homolog of, MADR2 antibody
    • hMAD-2 antibody
    • HsMAD2 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 decapentaplegic, Drosophila, homolog of, 2 antibody
    • Mothers against DPP homolog 2 antibody
    • OTTHUMP00000163489 antibody
    • Sma and Mad related protein 2 antibody
    • Sma- and Mad-related protein 2 MAD antibody
    • SMAD 2 antibody
    • SMAD family member 2 antibody
    • SMAD, mothers against DPP homolog 2 antibody
    • SMAD2 antibody
    • SMAD2_HUMAN antibody
    see all

Images

  • Overlay histogram showing PC3 cells stained with ab71109 (red line). The cells were fixed with 80% 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 (ab71109, 0.1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
  • All lanes : Anti-Smad2 antibody [7A5] (ab71109) at 1/2000 dilution

    Lane 1 : HeLa cell lysate
    Lane 2 : A431 cell lysate
    Lane 3 : NIH3T3 cell lysate
    Lane 4 : PC12 cell lysate


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

References

This product has been referenced in:
  • Pereira PN  et al. Antagonism of Nodal signaling by BMP/Smad5 prevents ectopic primitive streak formation in the mouse amnion. Development 139:3343-54 (2012). Read more (PubMed: 22912414) »

See 1 Publication for this product

Customer reviews and Q&As

Application
Western blot
Sample
Human Cell lysate - nuclear (Normal Human Keratinocyte)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
Normal Human Keratinocyte
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Apr 15 2015

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Mouse Cell lysate - whole cell (lung epithelium)
Total protein in input
600 µg
Specification
lung epithelium
Immuno-precipitation step
Protein A/G
Username

Abcam user community

Verified customer

Submitted May 18 2010

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

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