Anti-SMURF2 Polyclonal Antibody is a Rabbit antibody targeting SMURF2. Anti-SMURF2 Polyclonal Antibody can be used in WB,IHC-P,IHC-Fr,IF,FCM.

IgG

Human,Mouse (predicted:Rat,Chicken,Cow,Horse)

1. Paraformaldehyde-fixed, paraffin embedded (mouse brain tissue); Antigen retrieval by microwave in sodium citratebuffer (pH6.0); Block endogenous peroxidase by 3% hydrogen peroxide for 30 minutes; Blocking buffer (3%BSA) at RTfor 30 min; Antibody incubation with (SMURF2) Polyclonal/MonoclonalAntibody, Unconjugated (TMAB-12986) at 1:400 overnight at 4℃,followed by conjugation to the secondary antibody (labeled with HRP) and DAB staining.
2. Sample: Placenta (Mouse) Lysate at 40 μg
Primary: Anti-SMURF2 (TMAB-12986) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 82 kD
Observed band size: 82 kD
3. Blank control: A431.
Primary Antibody (green line): Rabbit Anti-SMURF2 antibody (TMAB-12986)
Dilution: 3 μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-AF647
Dilution: 3 μg /test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at-20℃. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:100-500; FCM: 3μg/Test

Polyclonal

KLH conjugated synthetic peptide: human SMURF2

Human

SMURF2

E3 ubiquitin-protein ligase SMURF2

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.