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    smurf-2-antibody-ep629y3-ab53316.pdf

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Epigenetics and Nuclear Signaling Transcription Other factors
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Validated using a knockout cell lineRecombinantRabMAb

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

  • Datasheet
Reviews (2) Submit a question References (15)

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Western blot - Anti-SMURF 2 antibody [EP629Y3] (ab53316)
  • Western blot - Anti-SMURF 2 antibody [EP629Y3] (ab53316)
  • Western blot - Anti-SMURF 2 antibody [EP629Y3] (ab53316)
  • Anti-SMURF 2 antibody [EP629Y3] (ab53316)

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: Human

Conjugates logo Related conjugates and formulations

Carrier Free

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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: Human
    Predicted to work with: Mouse, Rat
  • 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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EP629Y3
  • Isotype

    IgG
  • Research areas

    • Epigenetics and Nuclear Signaling
    • Transcription
    • Other factors
    • Stem Cells
    • Signaling Pathways
    • TGF beta
    • Cytoplasmic

Associated products

  • Alternative Versions

    • Anti-SMURF 2 antibody [EP629Y3] - BSA and Azide free (ab239855)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human SMURF2 knockout HeLa cell line (ab265231)
  • KO cell lysates

    • Human SMURF2 knockout HeLa cell lysate (ab257691)
  • Recombinant Protein

    • Recombinant human SMURF 2 protein (ab95930)

Applications

The Abpromise guarantee

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 (2)
1/500 - 1/1000. Detects a band of approximately 86 kDa (predicted molecular weight: 86 kDa).
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.
  • Target information above from: UniProt accession Q9HAU4 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 64750 Human
    • Entrez Gene: 66313 Mouse
    • Entrez Gene: 303614 Rat
    • Omim: 605532 Human
    • SwissProt: Q9HAU4 Human
    • SwissProt: A2A5Z6 Mouse
    • Unigene: 340955 Mouse
    • 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

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

    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

    Datasheets and documents

    • Datasheet download

      Download

    References (15)

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

    ab53316 has been referenced in 15 publications.

    • Wang Y  et al. Extracellular-vesicle containing miRNA-503-5p released by macrophages contributes to atherosclerosis. Aging (Albany NY) 13:12239-12257 (2021). PubMed: 33872218
    • Tang LY  et al. Phosphorylation of SMURF2 by ATM exerts a negative feedback control of DNA damage response. J Biol Chem 295:18485-18493 (2020). PubMed: 33097595
    • Cui Y  et al. CUEDC1 inhibits epithelial-mesenchymal transition via the TßRI/Smad signaling pathway and suppresses tumor progression in non-small cell lung cancer. Aging (Albany NY) 12:20047-20068 (2020). PubMed: 33099540
    • Wang R  et al. Synergistic effects of miR-708-5p and miR-708-3p accelerate the progression of osteoporosis. J Int Med Res 48:300060520978015 (2020). PubMed: 33322976
    • 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
    View all Publications for this product

    Customer reviews and Q&As

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

    1-2 of 2 Abreviews or Q&A

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

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Dopaminergic Neurons)
    Gel Running Conditions
    Reduced Denaturing (NuPage 4-12% MES buffer)
    Loading amount
    30 µg
    Specification
    Dopaminergic Neurons
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Nov 20 2020

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

    Inconclusive
    Abreviews
    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
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Dr. Frank Götschel

    Verified customer

    Submitted Sep 09 2009

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