Phospho-Smad2/Smad3 (Thr8) Rabbit Polyclonal Antibody

• Catalog Number: orb186235
• Category: Antibodies
• Description: Phospho-Smad2/Smad3 (Thr8) Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, WB
• Predicted Reactivity: Bovine, Canine, Equine, Gallus, Porcine, Rat
• Reactivity: Human, Mouse
• Isotype: IgG
• Immunogen: KLH conjugated synthesised phosphopeptide derived from human Smad2/Smad3 around the phosphorylation site of Thr8 PF(p-T)PP
• Antibody Type: Primary Antibody
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, Flow-Cyt=1μg/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 52 kDa
• Target: SMAD2
• UniProt ID: Q15796
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Alternative names: Smad2+Smad3(phospho T8); p-Smad2+Smad3(phospho T8)
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Blank control: Hela. Primary Antibody (green line): Rabbit Anti-phospho-Smad2/Smad3 (Thr8) antibody (orb186235), 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. Acquisition of 20000 events was performed.

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.

Sample: cerebrum (mouse) Lysate at 40 ug, RSC96 cell (rat) Lysate at 30 ug, hepG2 cell (human) Lysate at 30 ug, Hcclm3 cell (human) Lysate at 30 ug, Primary: Anti-p-Smad2/Smad3 (Thr8) (orb186235) at 1/500 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 52kD, Observed band size: 48, 52 kD.

Sample: HUVEC cell (human) Lysate at 30 ug, Primary: Anti-p-Smad2/Smad3 (Thr8) (orb186235) at 1/500 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 52kD, Observed band size: 52 kD.

Sample: U251 cell (human) Lysate at 30 ug, Primary: Anti-p-Smad2/Smad3 (Thr8) (orb186235) at 1/500 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 52kD, Observed band size: 52 kD.