Overview

  • Product name
    Anti-SMURF 2 antibody
    See all SMURF 2 primary antibodies
  • Description
    Rabbit polyclonal to SMURF 2
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat, Chicken, Cow, Dog, Zebrafish
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 200 - 300 of Human SMURF 2.

    Read Abcam's proprietary immunogen policy

    (Peptide available as ab109647.)

  • Positive control
    • This antibody gave a positive signal in the following whole cell lysates: HeLa; HepG2; Caco2; MCF7; SHSY5Y; U20S; A549.

Properties

Applications

Our Abpromise guarantee covers the use of ab94483 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
ICC/IF Use a concentration of 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 86 kDa (predicted molecular weight: 86 kDa).

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
    see all
  • 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 (ab94483)
    Western blot - Anti-SMURF 2 antibody (ab94483)
    Anti-SMURF 2 antibody (ab94483) at 1 µg/ml + U2OS (Human osteosarcoma cell line) Whole Cell Lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 86 kDa
    Observed band size: 86 kDa
    Additional bands at: 25 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 3 minutes
  • Immunocytochemistry/ Immunofluorescence - Anti-SMURF 2 antibody (ab94483)
    Immunocytochemistry/ Immunofluorescence - Anti-SMURF 2 antibody (ab94483)
    ICC/IF image of ab94483 stained HepG2 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab94483, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 100% methanol fixed (5 min) HepG2 cells at 5µg/ml.

References

ab94483 has not yet been referenced specifically in any publications.

Publishing research using ab94483? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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2 reviews or Q&A

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Western blot abreview for Anti-SMURF 2 antibody

 Good
Abreviews
Application
Western blot
Sample
Zebrafish Tissue lysate - whole (Brain)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
20 µg
Specification
Brain
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Read More
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Melek Umay Tüz

Verified customer

Submitted May 05 2015

Western blot abreview for Anti-SMURF 2 antibody

 Good
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Breast cell line)
Loading amount
100000 cells
Specification
Breast cell line
Gel Running Conditions
Reduced Denaturing (12.5%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Read More
abreview image
Username

Abcam user community

Verified customer

Submitted Nov 21 2012

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