SMAD7 Rabbit Polyclonal Antibody

• Catalog Number: orb500819
• Category: Antibodies
• Description: SMAD7 Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, ICC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Bovine, Porcine
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from human Smad7 (1-100/426aa)
• Antibody Type: Primary Antibody
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, ICC/IF=1:100, IF=1:100-500, Flow-Cyt=1ug/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 46 kDa
• Target: SMAD7
• UniProt ID: O15105
• 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: MAD(mothers against decapentaplegic Drosophila) ho
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Blank control: SH-SY5Y. Primary Antibody (green line): Rabbit Anti-MADH7/Smad7 antibody (orb500819), 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.

Paraformaldehyde-fixed, paraffin embedded (mouse 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 (MADH7) Polyclonal Antibody, Unconjugated (orb500819) 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 (MADH7) Polyclonal Antibody, Unconjugated (orb500819) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (rat lung), 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 (Smad7) Polyclonal Antibody, Unconjugated (orb500819) at 1:600 overnight at 4°C, followed by a conjugated secondary for 20 minutes and DAB staining.

Paraformaldehyde-fixed, paraffin embedded (rat stomach), 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 (MADH7) Polyclonal Antibody, Unconjugated (orb500819) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Sample: Lane 1: Cerebrum (Mouse) Lysate at 40 ug, Lane 2: Cerebrum (Rat) Lysate at 40 ug, Lane 3: Stomach (Mouse) Lysate at 40 ug, Lane 4: Lung (Mouse) Lysate at 40 ug, Lane 5: Kidney (Mouse) Lysate at 40 ug, Primary: Anti-MADH7/Smad7 (orb500819) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 50 kD, Observed band size: 50 kD.

Sample: Lane 1: Stomach (Mouse) Lysate at 40 ug, Lane 2: Spleen (Mouse) Lysate at 40 ug, Lane 3: Lung (Mouse) Lysate at 40 ug, Primary: Anti-MADH7/Smad7 (orb500819) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 50 kD, Observed band size: 50 kD.

Sample: Lung (Mouse) Lysate at 30 ug, Placenta (Mouse) Lysate at 30 ug, Large intestine (Mouse) Lysate at 30 ug, Primary: Anti-MADH7/Smad7 (orb500819) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 46 kD, Observed band size: 50 kD.

U-2OS 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 (MADH7/Smad7) polyclonal Antibody, Unconjugated (orb500819) 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.