Antibody Type: Recombinant Rabbit Monoclonal

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

Reactivity: Human,Mouse,Rat

IgG

4E515

Human,Mouse,Rat

1. Blank control: A431. Primary Antibody (green line): Rabbit Anti-Smad2 antibody (TMAB-01737)
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.
2. The Hela (H) 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 then were incubated in 5% BSA to block non-specific protein-protein interactions (30 min at room temperature). Primary Antibody (green): Rabbit Anti-Smad2 antibody (TMAB-01737): 1 μg/10^6 cells; Secondary Antibody (white blue): Goat anti-Rabbit IgG-FITC: 1 μg/test. Isotype Control (orange): Rabbit IgG. Blank control (black): PBS.
3. Paraformaldehyde-fixed, paraffin embedded Mouse Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Smad2 Monoclonal Antibody, Unconjugated (TMAB-01737) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.
4. Paraformaldehyde-fixed, paraffin embedded Rat Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Smad2 Monoclonal Antibody, Unconjugated (TMAB-01737) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.
5. Paraformaldehyde-fixed, paraffin embedded Human Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Smad2 Monoclonal Antibody, Unconjugated (TMAB-01737) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.
6. Paraformaldehyde-fixed, paraffin embedded Mouse Cerebellum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Smad2 Monoclonal Antibody, Unconjugated (TMAB-01737) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.
7. Paraformaldehyde-fixed, paraffin embedded Rat Cerebellum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Smad2 Monoclonal Antibody, Unconjugated (TMAB-01737) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.
8. 25 μg total protein per Lane of various lysates probed with Smad2 monoclonal antibody, unconjugated (TMAB-01737) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at RT for 60 min.
9. 25 μg total protein per Lane of various lysates probed with SMAD2 monoclonal antibody, unconjugated (TMAB-01737) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at RT for 60 min.

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

WB: 1:500-2000; IHC-P: 1:50-200; IHC-Fr: 1:50-200; ICC/IF: 1:50-200; IF: 1:50-200; FCM: 1ug/Test

Monoclonal

A synthesized peptide: human SMAD2

Human

4087

TGF,SMADs,SMADs,Cytoplasmic