Phospho-SMAD2 (Ser464) Rabbit Polyclonal Antibody

• Catalog Number: orb704513
• Category: Antibodies
• Description: Phospho-SMAD2 (Ser464) Rabbit Polyclonal Antibody
• Target: SMAD2
• Clonality: Polyclonal
• Species/Host: Rabbit
• Isotype: IgG
• Conjugation: Unconjugated
• Reactivity: Human, Mouse
• Predicted Reactivity: Bovine, Canine, Equine, Gallus, Rat
• Form/Appearance: Liquid
• Concentration: 1mg/ml
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Purification: Affinity purified by Protein A
• Immunogen: KLH conjugated synthesised phosphopeptide derived from human Smad2 around the phosphorylation site of Ser464 RC(p-S)SM
• UniProt ID: Q15796
• MW: 58 kDa
• Tested applications: FC, IF, IHC-Fr, IHC-P, WB
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, IF=1:100-500, Flow-Cyt=3ug/Test
• Antibody Type: Primary Antibody
• 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.
• Alternative names: phospho-Smad2(p-Ser464); p-Smad2(Ser464); phospho-
• Research Area: Cell Biology, Cell Signaling, Cytoplasmic, Intrace
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Blank control (Black line): Mouse spleen (Black). Primary Antibody (green line): Rabbit Anti-phospho-SMAD2 (Ser425) antibody, 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. Acquisition of 20000 events was performed.

Paraformaldehyde-fixed, paraffin embedded (human placenta), 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-SMAD2 (Ser464)) Polyclonal Antibody, Unconjugated (orb704513) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (mouse brain), 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-SMAD2 (Ser464)) Polyclonal Antibody, Unconjugated (orb704513) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Sample: Lane 1: Heart (Mouse) Lysate at 40 ug, Lane 2: Muscle (Mouse) Lysate at 40 ug, Primary: Anti-phospho-SMAD2 (Ser464) (orb704513) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 58 kD, Observed band size: 58 kD.

Sample: Lane 1: Hela (Human) Cell Lysate at 30 ug, Lane 2: HT1080 (Human) Cell Lysate at 30 ug, Lane 3: Jurkat (Human) Cell Lysate at 30 ug, Lane 4: A549 (Human) Cell Lysate at 30 ug, Lane 5: 293T (Human) Cell Lysate at 30 ug, Primary: Anti-phospho-SMAD2 (Ser464) (orb704513) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 60 kD, Observed band size: 60 kD.