Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-SMURF 2 antibody [EP629Y3] (ab53316)

Reviews (1) Submit a question References (11)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP629Y3] to SMURF 2
  • Suitable for: WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-SMURF 2 antibody [EP629Y3]
    See all SMURF 2 primary antibodies

  • Description

    Rabbit monoclonal [EP629Y3] to SMURF 2

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WBmore details
    Unsuitable for: Flow Cyt,ICC,IHC-P or IP

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human SMURF 2 aa 200-300. The exact sequence is proprietary.

  • Positive control

    • WB: HeLa, U-2 OS and SH-SY-5Y cell lysates.

  • 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.

Properties

Applications

Our Abpromise guarantee covers the use of ab53316 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 1/500 - 1/1000. Detects a band of approximately 86 kDa (predicted molecular weight: 86 kDa).
  • Application notes
    Is unsuitable for Flow Cyt,ICC,IHC-P or IP.

    • Target

    • Function

      E3 ubiquitin-protein ligase which accepts ubiquitin from an E2 ubiquitin-conjugating enzyme in the form of a thioester and then directly transfers the ubiquitin to targeted substrates. Interacts with SMAD1 and SMAD7 in order to trigger their ubiquitination and proteasome-dependent degradation. In addition, interaction with SMAD7 activates autocatalytic degradation, which is prevented by interaction with SCYE1. Forms a stable complex with the TGF-beta receptor-mediated phosphorylated SMAD2 and SMAD3. In this way, SMAD2 may recruit substrates, such as SNON, for ubiquitin-mediated degradation. Enhances the inhibitory activity of SMAD7 and reduces the transcriptional activity of SMAD2. Coexpression of SMURF2 with SMAD1 results in considerable decrease in steady-state level of SMAD1 protein and a smaller decrease of SMAD2 level.

    • Tissue specificity

      Widely expressed.

    • Pathway

      Protein modification; protein ubiquitination.

    • Sequence similarities

      Contains 1 C2 domain.
      Contains 1 HECT (E6AP-type E3 ubiquitin-protein ligase) domain.
      Contains 3 WW domains.

    • Domain

      The second and third WW domains are responsible for interaction with the PY-motif of R-SMAD (SMAD1, SMAD2 and SMAD3).
      The C2 domain is involved in autoinhibition of the catalytic activity by interacting with the HECT domain.

    • Post-translational
      modifications

      Auto-ubiquitinated and ubiquitinated in the presence of RNF11 and UBE2D1.

    • Cellular localization

      Nucleus. Cytoplasm. Cell membrane. Membrane raft. Cytoplasmic in the presence of SMAD7. Co-localizes with CAV1, SMAD7 and TGF-beta receptor in membrane rafts.
    • Information by UniProt

    • Database links

      • Alternative names

        • E3 ubiquitin-protein ligase SMURF2 antibody
        • EC 6.3.2. antibody
        • hSMURF2 antibody
        • MGC138150 antibody
        • Smad specific E3 ubiquitin ligase 2 antibody
        • SMAD specific E3 ubiquitin protein ligase 2 antibody
        • SMAD ubiquitination regulatory factor 2 antibody
        • SMAD-specific E3 ubiquitin-protein ligase 2 antibody
        • SMUF2_HUMAN antibody
        • Smurf2 antibody
        • Ubiquitin protein ligase SMURF2 antibody
        see all

      Images

      • Western blot - Anti-SMURF 2 antibody [EP629Y3] (ab53316)
        Western blot - Anti-SMURF 2 antibody [EP629Y3] (ab53316)
        All lanes : Anti-SMURF 2 antibody [EP629Y3] (ab53316) at 1/1000 dilution

        Lane 1 : Wild-type HeLa cell lysate
        Lane 2 : SMURF2 knockout HeLa cell lysate
        Lane 3 : SH-SY5Y cell lysate

        Lysates/proteins at 20 µg per lane.

        Secondary
        All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

        Predicted band size: 86 kDa
        Observed band size: 86 kDa



        Lanes 1-3: Merged signal (red and green). Green - ab53316 observed at 86 kDa. Red - loading control ab8245 observed at 36 kDa.

         ab53316 Anti-SMURF 2 antibody [EP629Y3]  was shown to specifically react with SMURF 2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265231 (knockout cell lysate ab257691) was used. Wild-type and SMURF 2 knockout samples were subjected to SDS-PAGE. ab53316 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

      • Western blot - Anti-SMURF 2 antibody [EP629Y3] (ab53316)
        Western blot - Anti-SMURF 2 antibody [EP629Y3] (ab53316)
        All lanes : Anti-SMURF 2 antibody [EP629Y3] (ab53316) at 1/1000 dilution

        Lane 1 : Wild-type HeLa cell lysate
        Lane 2 : SMURF2 knockout HeLa cell lysate
        Lane 3 : U-2 OS cell lysate

        Lysates/proteins at 20 µg per lane.

        Secondary
        All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

        Predicted band size: 86 kDa
        Observed band size: 86 kDa



        Lanes 1-3: Merged signal (red and green). Green - ab53316 observed at 86 kDa. Red - loading control ab8245 observed at 36 kDa. 

         ab53316 Anti-SMURF 2 antibody [EP629Y3] was shown to specifically react with SMURF 2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264806 (knockout cell lysate ab257690) was used.  Wild-type and SMURF 2 knockout samples were subjected to SDS-PAGE.  ab53316 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

      • Western blot - Anti-SMURF 2 antibody [EP629Y3] (ab53316)
        Western blot - Anti-SMURF 2 antibody [EP629Y3] (ab53316)
        Anti-SMURF 2 antibody [EP629Y3] (ab53316) at 1/500 dilution + SH-SY-5Y cell lysate at 10 µg

        Secondary
        Goat anti-rabbit HRP labeled at 1/2000 dilution

        Predicted band size: 86 kDa
        Observed band size: 86 kDa

      Protocols

      To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

      Click here to view the general protocols

      References (11)

      Publishing research using ab53316? Please let us know so that we can cite the reference in this datasheet.

      ab53316 has been referenced in 11 publications.

      • Mund T & Pelham HR Substrate clustering potently regulates the activity of WW-HECT domain-containing ubiquitin ligases. J Biol Chem 293:5200-5209 (2018). PubMed: 29463679
      • Huang PS  et al. The Long Non-Coding RNA MIR503HG Enhances Proliferation of Human ALK-Negative Anaplastic Large-Cell Lymphoma. Int J Mol Sci 19:N/A (2018). WB . PubMed: 29758012
      • Cui X  et al. Baicalein represses TGF-ß1-induced fibroblast differentiation through the inhibition of miR-21. Toxicol Appl Pharmacol 358:35-42 (2018). PubMed: 30201452
      • Mund T  et al. Ubiquitination of alpha-synuclein filaments by Nedd4 ligases. PLoS One 13:e0200763 (2018). PubMed: 30021006
      • Wang J  et al. MicroRNA-485 Modulates the TGF-ß/ Smads Signaling Pathway in Chronic Asthmatic Mice by Targeting Smurf2. Cell Physiol Biochem 51:692-710 (2018). PubMed: 30463065
      View all Publications for this product

      Customer reviews and Q&As

      Show All Reviews Q&A
      Submit a review Submit a question

      Western blot abreview for Anti-SMURF 2 antibody [EP629Y3]

       Inconclusive
      Abreviews
      abreview image
      Application
      Western blot
      Sample
      Mouse Cell lysate - whole cell (mouse liver cell line)
      Loading amount
      15 µg
      Specification
      mouse liver cell line
      Gel Running Conditions
      Reduced Denaturing (8%)
      Blocking step
      Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
      Read More

      Dr. Frank Götschel

      Verified customer

      Submitted Sep 09 2009

      Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
      For licensing inquiries, please contact partnerships@abcam.com