Anti-Phospho-Smad2/3 (Thr8) Polyclonal Antibody is a Rabbit antibody targeting Phospho-Smad2/3 (Thr8). Anti-Phospho-Smad2/3 (Thr8) Polyclonal Antibody can be used in FCM,WB.

IgG

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

1. Blank control: Hela.
Primary Antibody (green line): Rabbit Anti-phospho-Smad2/Smad3 (Thr8) antibody (TMAB-01497)
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. blank: A549 cells (blue line)
isotype control: rabbit IgG (orange line)
second antibody: goat anti-rabbit IgG (white blue line)
primary antibody: rabbit Anti-phospho-Smad2/Smad3 (Thr8) (green line); contration: 3 μg/10^6 cells
3. Sample:
HUVEC cell (human) Lysate at 30 μg
Primary: Anti-p-Smad2/Smad3 (Thr8) (TMAB-01497) at 1/500 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 52 kDa
Observed band size: 52 kDa
4. Sample:
cerebrum (mouse) Lysate at 40 μg
RSC96 cell (rat) Lysate at 30 μg
hepG2 cell (human) Lysate at 30 μg
Hcclm3 cell (human) Lysate at 30 μg
Primary: Anti-p-Smad2/Smad3 (Thr8) (TMAB-01497) at 1/500 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 52 kDa
Observed band size: 48,52 kDa
5. Sample:
U251 cell (human) Lysate at 30 μg
Primary: Anti-p-Smad2/Smad3 (Thr8) (TMAB-01497) at 1/500 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 52 kDa
Observed band size: 52 kDa

WB: 1:500-2000; FCM: 1μg/Test

Polyclonal

KLH conjugated synthesised phosphopeptide: human Smad2/Smad3 around the phosphorylation site of Thr8

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.