Recombinant
RabMAb

Recombinant Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334)

Overview

  • Product name

    Anti-Smad2 (phospho S255) antibody [EPR2856(N)]
    See all Smad2 primary antibodies
  • Description

    Rabbit monoclonal [EPR2856(N)] to Smad2 (phospho S255)
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Dot blot, IHC-P, IP, WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human Smad2 aa 200-300 (phospho S255). The exact sequence is proprietary.
    Database link: Q15796

  • Positive control

    • Hela treated with Okadaic acid and Calyculin A, Hela treated with Okadaic acid and Calyculin A, Human endometrium, Human transitional cell carcinoma of bladder.
  • General notes

     

     

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab188334 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
Dot blot Use at an assay dependent concentration.
IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP 1/50 - 1/70.
WB 1/1000 - 1/10000. Detects a band of approximately 58 kDa (predicted molecular weight: 52 kDa).Can be blocked with Human Smad2 (phospho S255) peptide (ab225544).

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

  • All lanes : Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334) at 1/10000 dilution

    Lane 1 : Hela treated with Okadaic acid and Calyculin A
    Lane 2 : Hela treated with Okadaic acid and Calyculin A , then treated with Lambda phosphorylase
    Lane 3 : Untreated Hela

    Lysates/proteins at 10 µg per lane.

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

    Predicted band size: 52 kDa

  • Immunohistochemical analysis of formalin fixed paraffin embedded Human endometrium labeling Smad2 (phospho S255) with ab188334 at 1/100 dilution and HRP Polymer for Rabbit IgG. Counterstained with Hematoxylin.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of formalin fixed paraffin embedded Human transitional cell carcinoma of bladder labeling Smad2 (phospho S255) with ab188334 at 1/100 dilution and HRP Polymer for Rabbit IgG.  Counterstained with Hematoxylin.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Dot blot analysis of Smad2 (S255) phospho peptide (Lane 1), Smad2 non-phospho peptide (Lane 2), labelling Smad2 (S255) phospho peptide with ab188334 at a dilution of 1:1000 dilution (1.365ug/ml). A Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) was used as the secondary antibody at a dilution of 1:20,000 dilution. Blocking buffer: 5% NFDM/TBST. Dilution buffer: 5% NFDM /TBST .

  • Immunoprecipitation of Hela cells treated with Okadaic acis and Calyculin A (Lane 1) or PBS (Lane 2) labeling Smad2 (phospho S255) with ab188334 at 1/50 dilution and Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1500 dilution

References

This product has been referenced in:

  • Huo Q  et al. Acetazolamide attenuates cardiac fibrosis induced by aortic constriction through inhibiting transforming growth factor-ß1/Smad2 signaling pathway in mice. Exp Ther Med 17:2317-2321 (2019). Read more (PubMed: 30867716) »
  • Dong R  et al. Bushenhuoxue Formula Facilitates Articular Cartilage Repair and Attenuates Matrix Degradation by Activation of TGF-ß Signaling Pathway. Evid Based Complement Alternat Med 2018:2734581 (2018). Read more (PubMed: 30402118) »
See all 12 Publications for this product

Customer reviews and Q&As

Application
Immunocytochemistry/ Immunofluorescence
Sample
Hamster Cell (ovary)
Permeabilization
No
Specification
ovary
Blocking step
goat serum/0.2% triton X100 as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Paraformaldehyde

Prof. Shyamal Roy

Verified customer

Submitted Jun 02 2016

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