The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/500 - 1/1000. Detects a band of approximately 86 kDa (predicted molecular weight: 86 kDa).
Is unsuitable for Flow Cyt,ICC,IHC-P or IP.
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.
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.
Auto-ubiquitinated and ubiquitinated in the presence of RNF11 and UBE2D1.
Nucleus. Cytoplasm. Cell membrane. Membrane raft. Cytoplasmic in the presence of SMAD7. Co-localizes with CAV1, SMAD7 and TGF-beta receptor in membrane rafts.
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
This product has been referenced in:
Mund T & Pelham HR Substrate clustering potently regulates the activity of WW-HECT domain-containing ubiquitin ligases. J Biol Chem293:5200-5209 (2018).
Read more (PubMed: 29463679) »
Maxfield KE et al. Comprehensive functional characterization of cancer-testis antigens defines obligate participation in multiple hallmarks of cancer. Nat Commun6:8840 (2015).
Read more (PubMed: 26567849) »