Anti-SMURF2 antibody (bs-4056R)

Anti-SMURF2 antibody 

DATASHEET

Host:Rabbit

Target Protein:SMURF2

IR:Immunogen Range:601-700/748

Clonality:Polyclonal

Isotype:IgG

Entrez Gene:64750

Swiss Prot:Q9HAU4

Source:KLH conjugated synthetic peptide derived from human SMURF2:601-700/748 

Purification:affinity purified by Protein A

Storage:0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol. Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.

Background:SMURF2, a 748-amino acid ubiquitin E3 ligase that is 83% identical to SMURF1, codes for a C2-WW-HECT domain ubiquitin ligase that associates constitutively with SMAD7. Binding to SMAD7 induces export of SMURF2 and recruitment to the activated transforming growth factor-beta receptor (TGFBR), where it causes receptor and SMAD7 degradation. A strong interaction of second and third SMURF2 WW domains has been identified with SMAD1, SMAD2, and SMAD3, but not SMAD4. Western blot analysis showed that SMURF2 selectively downregulates the transcription of SMAD2 and SMAD1, but not SMAD3. The nuclear SMURF2/phosphorylated SMAD2 interaction is requires TGFB1.

Size:100ul

Concentration:1mg/ml

Applications:WB(1:500-2000)
ELISA(1:5000-10000)
IHC-P(1:100-500)
IHC-F(1:100-500)
Flow-Cyt(3ug/test)
IF(1:100-500)

Cross Reactive Species:Human
Mouse
Rat
Chicken
Cow
Horse
.

For research use only. Not intended for diagnostic or therapeutic use.

VALIDATION IMAGES

Sample: Placenta (Mouse) Lysate at 40 ug
Primary: Anti-SMURF2 (bs-4056R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 82 kD
Observed band size: 82 kD

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 30min; Antibody incubation with (SMURF2) Polyclonal/MonoclonalAntibody, Unconjugated (bs-4056R) at 1:400 overnight at 4℃,followed by conjugation to the secondary antibody (labeled with HRP)and DAB staining.

Blank control:A431.
Primary Antibody (green line): Rabbit Anti-SMURF2 antibody (bs-4056R)
Dilution: 3μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-AF647
Dilution: 3μg /test.
Protocol
The cells were fixed with 4% PFA (10min 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.