Recombinant
RabMAb

Recombinant Anti-Smad2 (phospho S255) antibody [EPR2856(N)] - BSA and Azide free (ab219598)

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Overview

  • Product name

    Anti-Smad2 (phospho S255) antibody [EPR2856(N)] - BSA and Azide free
    See all Smad2 primary antibodies

  • Description

    Rabbit monoclonal [EPR2856(N)] to Smad2 (phospho S255) - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IP, WB, IHC-P, Dot blotmore 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

    Ab219598 is the carrier-free version of ab188334. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab219598 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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 ab219598 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
IP Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 58 kDa (predicted molecular weight: 52 kDa).
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Dot blot Use at an assay dependent concentration.

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad2 (phospho S255) antibody [EPR2856(N)] - BSA and Azide free (ab219598)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad2 (phospho S255) antibody [EPR2856(N)] - BSA and Azide free (ab219598)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab188334).

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

  • Immunoprecipitation - Anti-Smad2 (phospho S255) antibody [EPR2856(N)] - BSA and Azide free (ab219598)
    Immunoprecipitation - Anti-Smad2 (phospho S255) antibody [EPR2856(N)] - BSA and Azide free (ab219598)

    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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab188334).

  • Dot Blot - Anti-Smad2 (phospho S255) antibody [EPR2856(N)] - BSA and Azide free (ab219598)
    Dot Blot - Anti-Smad2 (phospho S255) antibody [EPR2856(N)] - BSA and Azide free (ab219598)

    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 .

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab188334).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad2 (phospho S255) antibody [EPR2856(N)] - BSA and Azide free (ab219598)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad2 (phospho S255) antibody [EPR2856(N)] - BSA and Azide free (ab219598)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab188334).

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

References

ab219598 has not yet been referenced specifically in any publications.

Publishing research using ab219598? Please let us know so that we can cite the reference in this datasheet.

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