Antibody Type: Rabbit Polyclonal

Application: WB,IHC-P,IHC-Fr,IF,FCM

Reactivity: Human,Mouse,Rat (predicted:Chicken,Dog,Cow,Rabbit)

IgG

Human,Mouse,Rat (predicted:Chicken,Dog,Cow,Rabbit)

1. Tissue/cell: rat skin 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-Smad2 Polyclonal Antibody, Unconjugated (TMAB-01736) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
2. Tissue/cell: rat choroid 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-Smad2 Polyclonal Antibody, Unconjugated (TMAB-01736) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
3. Sample:
Lane 1: Cerebrum (Mouse) Lysate at 40 μg
Lane 2: Heart (Mouse) Lysate at 40 μg
Lane 3: Testis (Mouse) Lysate at 40 μg
Lane 4: Kidney (Mouse) Lysate at 40 μg
Lane 5: Placenta (Mouse) Lysate at 40 μg
Lane 6: Cerebrum (Rat) Lysate at 40 μg
Lane 7: Heart (Rat) Lysate at 40 μg
Lane 8: Testis (Rat) Lysate at 40 μg
Lane 9: Kidney (Rat) Lysate at 40 μg
Lane 10: Hela (Human) Cell Lysate at 30 μg
Lane 11: HT1080 (Human) Cell Lysate at 30 μg
Lane 12: Jurkat (Human) Cell Lysate at 30 μg
Lane 13: Raw264.7 (Mouse) Cell Lysate at 30 μg
Lane 14: HL60 (Human) Cell Lysate at 30 μg
Primary: Anti-Smad2 (TMAB-01736) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 60 kDa
Observed band size: 60 kDa
4. 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 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Smad2) Polyclonal Antibody, Unconjugated (TMAB-01736) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
5. Blank control (Black line): Mouse spleen (Black).
Primary Antibody (green line): Rabbit Anti-Smad2 antibody (TMAB-01736)
Dilution: 1 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.

WB,IHC-P,IHC-Fr,IF,FCM

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

Polyclonal

KLH conjugated synthetic peptide: human Smad2

Human

4087

TGF,SMADs,SMADs,Cytoplasmic