Smad3 Recombinant Rabbit Monoclonal Antibody

• Catalog Number: orb500265
• Category: Antibodies
• Description: Smad3 Recombinant Rabbit Monoclonal Antibody
• Target: SMAD3
• Clonality: Recombinant
• Species/Host: Rabbit
• Isotype: IgG
• Conjugation: Unconjugated
• Reactivity: Human, Mouse, Rat
• Predicted Reactivity: Mouse, Rat
• Form/Appearance: Liquid
• Concentration: 1mg/1ml
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Immunogen: Recombinant human Smad3 protein (1-200aa)
• UniProt ID: P84022
• MW: 47 kDa
• Tested applications: FC, ICC, IF, IHC-Fr, IHC-P, WB
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, ICC/IF=1:50-200, IF=1:100-500, Flow-Cyt=1ug/Test
• Antibody Type: Recombinant Antibody
• Clone Number: 3D1
• 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: hMAD 3; hSMAD3; HSPC193; JV15 2; JV152; MAD(mother
• Note: For research use only

Blank control: HUVEC. Primary Antibody (green line): Rabbit Anti-Smad3 antibody (orb500265), dilution: 1 ug/Test, Secondary Antibody: Goat anti-rabbit IgG-FITC, dilution: 0.5 ug/Test. Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 0.1% PBST for 20 min at room temperature. 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 (mouse liver), 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 (Smad3) Polyclonal Antibody, Unconjugated (orb500265) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (mouse ovary), 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 (Smad3) Polyclonal Antibody, Unconjugated (orb500265) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (rat 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 (Smad3) Polyclonal Antibody, Unconjugated (orb500265) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Raw264.7 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 (Smad3) monoclonal Antibody, Unconjugated (orb500265) 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.

Sample: A549 (Human) Cell Lysate at 30 ug, MG63 (Human) Cell Lysate at 30 ug, A431 (Human) Cell Lysate at 30 ug, Siha (Human) Cell Lysate at 30 ug, Primary: Anti-Smad3 (orb500265) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 52 kD, Observed band size: 52 kD.

Sample: Cerebrum (Mouse) Lysate at 40 ug, Cerebrum (Rat) Lysate at 40 ug, Large intestine (Mouse) Lysate at 40 ug, Ovary (Mouse) Lysate at 40 ug, Lung (Mouse) Lysate at 40 ug, Primary: Anti-Smad3 (orb500265) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 52 kD, Observed band size: 52 kD.