Smad2 Recombinant Rabbit Monoclonal Antibody

• Catalog Number: orb544569
• Category: Antibodies
• Description: Smad2 Recombinant Rabbit Monoclonal Antibody
• Target: SMAD2
• Clonality: Recombinant
• Species/Host: Rabbit
• Isotype: IgG
• Conjugation: Unconjugated
• Reactivity: Human, Mouse, Rat
• Predicted Reactivity: Mouse, 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: A synthesized peptide derived from human SMAD2 (230-270aa)
• UniProt ID: Q15796
• MW: 52 kDa
• Tested applications: FC, ICC, IF, IHC-Fr, IHC-P, WB
• Dilution range: WB=1:500-2000, IHC-P=1:50-200, IHC-F=1:50-200, ICC/IF=1:50-200, IF=1:50-200, Flow-Cyt=1ug/Test
• Antibody Type: Primary Antibody
• Clone Number: 9A3
• 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: Mothers against decapentaplegic homolog 2; SMAD 2;
• Research Area: Cell Biology, Cell Signaling, Cytoplasmic, Intrace
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

A549 cell, 4% Paraformaldehyde-fixed, Triton X-100 at room temperature for 20 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Antibody incubation with (Smad2) monoclonal Antibody, Unconjugated (orb544569) 1:100, 90 minutes at 37°C, followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.

Blank control: A431. Primary Antibody (green line): Rabbit Anti-Smad2 antibody (orb544569), dilution: 1 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-AF488, 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 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 (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 (Smad2) Polyclonal Antibody, Unconjugated (orb544569) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (mouse cerebellum), 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 (Smad2) Polyclonal Antibody, Unconjugated (orb544569) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (rat kidney), 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 (Smad2) Polyclonal Antibody, Unconjugated (orb544569) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Sample: Testis (Mouse) Lysate at 40 ug, A431 (Human) Cell Lysate at 30 ug, HepG2 (Human) Cell Lysate at 30 ug, HL60 (Human) Cell Lysate at 30 ug, Jurkat (Human) Cell Lysate at 30 ug, Primary: Anti-Smad2 (orb544569) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 52 kD, Observed band size: 53 kD.