Anti-Phospho-Smad1 (Ser463) Polyclonal Antibody is a Rabbit antibody targeting Phospho-Smad1 (Ser463). Anti-Phospho-Smad1 (Ser463) Polyclonal Antibody can be used in WB,IHC-P,IHC-Fr,IF,FCM.

IgG

Human,Mouse,Rat (predicted:Pig)

1. Sample: Muscle (Mouse) Lysate at 40 μg
Primary: Anti-Phospho-Smad1 (Ser463) (TMAB-11289) at 1/500 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 51 kD
Observed band size: 51 kD
2. Paraformaldehyde-fixed, paraffin embedded (human brain glioma); 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 (Smad1 (Ser463)) Polyclonal/MonoclonalAntibody, Unconjugated (TMAB-11289) at 1:400 overnight at 4℃,followed by conjugation to the secondary antibody (labeled with HRP) and DAB staining.
3. Paraformaldehyde-fixed, paraffin embedded (Rat heart); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Phospho-Smad1 (Ser463)) Polyclonal Antibody, Unconjugated (TMAB-11289) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
4. Sample: Heart (Mouse) Lysate at 40 μg
Primary: Anti-Phospho-Smad1 (Ser463) (TMAB-11289) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 51 kD
Observed band size: 51 kD
5. Blank control (black line): Hela.
Primary Antibody (green line): Rabbit Anti-Phospho-Smad1 (Ser463) antibody (TMAB-11289)
Dilution: 2 μg/Test;
Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC
Dilution: 0.5 μg/Test.
Isotype control (orange line): Normal Rabbit IgG
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 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: 2μg/Test

Polyclonal

KLH conjugated Synthesised phosphopeptide: human Smad1 around the phosphorylation site of Ser463

Human

SMAD1

Mothers against decapentaplegic homolog 1

Transcriptional modulator activated by BMP (bone morphogenetic proteins) type 1 receptor kinase. SMAD1 is a receptor-regulated SMAD (R-SMAD). SMAD1/OAZ1/PSMB4 complex mediates the degradation of the CREBBP/EP300 repressor SNIP1.