Anti-Phospho-Smad2 (Ser465) Polyclonal Antibody is a Rabbit antibody targeting Phospho-Smad2 (Ser465). Anti-Phospho-Smad2 (Ser465) Polyclonal Antibody can be used in WB,FCM.

IgG

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

1. Blank control: Hela.
Primary Antibody (green line): Rabbit Anti-phospho-Smad2 (Ser465) antibody (TMAB-01495)
Dilution: 1 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-AF647
Dilution: 1 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°C. 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.
2. Sample:
Lane 1: Hela (Human) Cell Lysate at 30 μg
Lane 2: HT1080 (Human) Cell Lysate at 30 μg
Lane 3: Jurkat (Human) Cell Lysate at 30 μg
Lane 4: HL60 (Human) Cell Lysate at 30 μg
Primary: Anti-phospho-Smad2 (Ser465) (TMAB-01495) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 60 kDa
Observed band size: 60 kDa
3. Blank control (Black line): Raji (Black).
Primary Antibody (green line): Rabbit Anti-phospho-Smad2 (Ser465) antibody (TMAB-01495)
Dilution: 3 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody (white blue line): Goat anti-rabbit IgG-PE
Dilution: 1 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 room temperature. The cells were then incubated in 5% BSA goat serum to block non-specific protein-protein interactions for 15 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.

WB: 1:500-5000; FCM: 1μg/Test

Polyclonal

KLH conjugated Synthesised phosphopeptide: human Smad2 around the phosphorylation site of Ser465

Human

SMAD2

Mothers against decapentaplegic homolog 2

TGF,SMADs,SMADs,Cytoplasmic

Receptor-regulated SMAD (R-SMAD) that is an intracellular signal transducer and transcriptional modulator activated by TGF-beta (transforming growth factor) and activin type 1 receptor kinases. Binds the TRE element in the promoter region of many genes that are regulated by TGF-beta and, on formation of the SMAD2/SMAD4 complex, activates transcription. May act as a tumor suppressor in colorectal carcinoma. Positively regulates PDPK1 kinase activity by stimulating its dissociation from the 14-3-3 protein YWHAQ which acts as a negative regulator.