RecombinantRabMAb

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

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR2856(N)] to Smad2 (phospho S255)
  • Suitable for: Dot blot, IHC-P, IP, WB
  • Reacts with: Mouse, Rat, Human

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

     

     

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

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).

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

    see all

  • 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

  • Western blot - Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334)
    Western blot - Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334)
    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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334)

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334)

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

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

    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

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

References (24)

Publishing research using ab188334? Please let us know so that we can cite the reference in this datasheet.

ab188334 has been referenced in 24 publications.

  • Lv Q  et al. Pirfenidone alleviates pulmonary fibrosis in vitro and in vivo through regulating Wnt/GSK-3ß/ß-catenin and TGF-ß1/Smad2/3 signaling pathways. Mol Med 26:49 (2020). PubMed: 32448163
  • Kozmikova I & Kozmik Z Wnt/ß-catenin signaling is an evolutionarily conserved determinant of chordate dorsal organizer. Elife 9:N/A (2020). PubMed: 32452768
  • Dhall S  et al. A Flowable Placental Formulation Prevents Bleomycin-Induced Dermal Fibrosis in Aged Mice. Int J Mol Sci 21:N/A (2020). PubMed: 32545915
  • Ying J  et al. Amygdalin Promotes Fracture Healing through TGF-ß/Smad Signaling in Mesenchymal Stem Cells. Stem Cells Int 2020:8811963 (2020). PubMed: 32963548
  • 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). PubMed: 30867716
View all Publications for this product

Customer reviews and Q&As

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1-2 of 2 Abreviews or Q&A

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Smad2 (phospho S255) antibody [EPR2856(N)]

 Excellent
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (NSCLC: A549 cells)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: pH6
Permeabilization
No
Specification
NSCLC: A549 cells
Blocking step
(agent) for 10 minute(s) · Concentration: 100% · Temperature: 22°C
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

Submitted Nov 24 2020

Immunocytochemistry/ Immunofluorescence abreview for Anti-Smad2 (phospho S255) antibody [EPR2856(N)]

 Excellent
Abreviews
abreview image
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
Read More

Prof. Shyamal Roy

Verified customer

Submitted Jun 02 2016

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