Phospho-Smad1 (Ser463) Rabbit Polyclonal Antibody

• Catalog Number: orb158426
• Category: Antibodies
• Description: Phospho-Smad1 (Ser463) Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Porcine
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated Synthesised phosphopeptide derived from human Smad1 around the phosphorylation site of Ser463 IS(p-S)VS
• Antibody Type: Primary Antibody
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, IF=1:100-500, Flow-Cyt=2ug/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 51 kDa
• Target: SMAD1
• UniProt ID: Q15797
• 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: Smad1(phospho S463); Smad1(phospho Ser463); p-Smad
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Blank control (black line): Hela. Primary Antibody (green line): Rabbit Anti-Phospho-Smad1 (Ser463) antibody (orb158426), dilution: 2 ug/Test, Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC, dilution: 0.5 ug/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°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.

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 (orb158426) at 1:400 overnight at 4°C, followed by conjugation to the secondary antibody (labeled with HRP) and DAB staining.

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 (orb158426) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.

Sample: Heart (Mouse) Lysate at 40 ug, Primary: Anti-Phospho-Smad1 (Ser463) (orb158426) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 51 kD, Observed band size: 51 kD.

Sample: Muscle (Mouse) Lysate at 40 ug, Primary: Anti-Phospho-Smad1 (Ser463) (orb158426) at 1/500 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 51 kD, Observed band size: 51 kD.