Anti-SMAD7 Polyclonal Antibody is a Rabbit antibody targeting SMAD7. Anti-SMAD7 Polyclonal Antibody can be used in WB,IHC-P,IHC-Fr,ICC/IF,IF,FCM.

IgG

Human,Mouse,Rat (predicted:Pig,Cow)

1. Tissue/cell: rat kidney tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37°C for 20 min;
Incubation: Anti-Smad7/Smad6 Polyclonal Antibody, Unconjugated 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
2. Sample:
Lung (Mouse) Lysate at 30 μg
Placenta (Mouse) Lysate at 30 μg
Large intestine (Mouse) Lysate at 30 μg
Primary: Anti-MADH7/Smad7 (TMAB-01742) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 46 kDa
Observed band size: 50 kDa
3. 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 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Smad7) Polyclonal Antibody, Unconjugated (TMAB-01742) at 1:600 overnight at 4°C, followed by a conjugated secondary for 20 min and DAB staining.
4. Blank control: SH-SY5Y.
Primary Antibody (green line): Rabbit Anti-MADH7/Smad7 antibody (TMAB-01742)
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.
5. Sample:
Lane 1: Cerebrum (Mouse) Lysate at 40 μg
Lane 2: Cerebrum (Rat) Lysate at 40 μg
Lane 3: Stomach (Mouse) Lysate at 40 μg
Lane 4: Lung (Mouse) Lysate at 40 μg
Lane 5: Kidney (Mouse) Lysate at 40 μg
Primary: Anti-MADH7/Smad7 (TMAB-01742) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 50 kDa
Observed band size: 50 kDa
6. 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 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (MADH7) Polyclonal Antibody, Unconjugated (TMAB-01742) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
7. 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 (TMAB-01742) 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 nucleus.
8. Sample:
Lane 1: Stomach (Mouse) Lysate at 40 μg
Lane 2: Spleen (Mouse) Lysate at 40 μg
Lane 3: Lung (Mouse) Lysate at 40 μg
Primary: Anti-MADH7/Smad7 (TMAB-01742) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 50 kDa
Observed band size: 50 kDa
9. 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 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (MADH7) Polyclonal Antibody, Unconjugated (TMAB-01742) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
10. 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 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (MADH7) Polyclonal Antibody, Unconjugated (TMAB-01742) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; ICC/IF: 1:100; IF: 1:100-500; FCM: 1ug/Test

Polyclonal

KLH conjugated synthetic peptide: human Smad7

Human

SMAD7

Mothers against decapentaplegic homolog 7

SMADs,SMADs

Antagonist of signaling by TGF-beta (transforming growth factor) type 1 receptor superfamily members; has been shown to inhibit TGF-beta (Transforming growth factor) and activin signaling by associating with their receptors thus preventing SMAD2 access. Functions as an adapter to recruit SMURF2 to the TGF-beta receptor complex. Also acts by recruiting the PPP1R15A-PP1 complex to TGFBR1, which promotes its dephosphorylation. Positively regulates PDPK1 kinase activity by stimulating its dissociation from the 14-3-3 protein YWHAQ which acts as a negative regulator.