Recombinant
RabMAb

Recombinant Anti-Smad2 antibody [EP567Y] - BSA and Azide free (ab216454)

Overview

  • Product name

    Anti-Smad2 antibody [EP567Y] - BSA and Azide free
    See all Smad2 primary antibodies
  • Description

    Rabbit monoclonal [EP567Y] to Smad2 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, Flow Cyt, ICC/IFmore details
    Unsuitable for: IHC-P or IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Smad2 aa 200-300.

  • Positive control

    • Jurkat cell lysate and human adenocarcinoma of uterus
  • General notes

    Ab216454 is the carrier-free version of ab33875. 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.

    ab216454 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 ab216454 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
WB Use at an assay dependent concentration. Detects a band of approximately 58 kDa (predicted molecular weight: 58 kDa).
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

ICC/IF Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for IHC-P or IP.
  • 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 Jurkat cells stained with purified ab33875 (pink line) at a dilution of 1/110. The cells were fixed with 2% PFA. FITC goat anti-rabbit was used at a dilution of 1/150 and rabbit monoclonal IgG was used as the isotype control (green line).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab33875).

    • Immunofluorescent staining of A673 cells, fixed with 4% PFA, using purified ab33875 at a dilution of 1/300. An Alexa Fluor® 555 goat anti-rabbit was used at 1/200. The negative control is shown in the bottom right hand panel - for the negative control, purified ab33875 was used at a dilution of 1/200 followed by an Alexa Fluor® 555 goat anti-mouse antibody at a dilution of 1/500.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab33875).

    • Overlay histogram showing PC3 cells stained with unpurified ab33875 (red line). The cells were fixed with 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 (ab33875, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in PC3 cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab33875).

    References

    This product has been referenced in:

    • Chen RY  et al. Effect of oridonin-mediated hallmark changes on inflammatory pathways in human pancreatic cancer (BxPC-3) cells. World J Gastroenterol 20:14895-903 (2014). WB ; Human . Read more (PubMed: 25356049) »
    • Gallo EM  et al. Angiotensin II-dependent TGF-ß signaling contributes to Loeys-Dietz syndrome vascular pathogenesis. J Clin Invest 124:448-60 (2014). IF . Read more (PubMed: 24355923) »
    See all 6 Publications for this product

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