Anti-Phospho-Smad2 (Thr220) Polyclonal Antibody 2 is a Rabbit antibody targeting Phospho-Smad2 (Thr220). Anti-Phospho-Smad2 (Thr220) Polyclonal Antibody 2 can be used in IHC-P,IHC-Fr,IF,FCM.

IgG

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

1. Tissue/cell: human placenta tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01 M, pH 6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37℃ for 20 min;
Incubation: Anti-Phospho-Smad2 (Thr220) Polyclonal Antibody, Unconjugated (TMAB-11298) 1: 200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining
2. Blank control: Hela.
Primary Antibody (green line): Rabbit Anti-Phospho-Smad2 (Thr220) antibody (TMAB-11298)
Dilution: 2 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℃. 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.

IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:100-500; FCM: 2μg/Test

Polyclonal

KLH conjugated Synthesised phosphopeptide: human Smad2 around the phosphorylation site of Thr220

Human

SMAD2

Mothers against decapentaplegic homolog 2

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.