Anti-Phospho-SMAD5 (Ser463+Ser465) Antibody (9Z247) is a Rabbit antibody targeting Phospho-SMAD5 (Ser463+Ser465). Anti-Phospho-SMAD5 (Ser463+Ser465) Antibody (9Z247) can be used in WB,IHC-P,IHC-Fr,ICC/IF,IF.

IgG

9Z247

Human,Mouse,Rat

1. Paraformaldehyde-fixed, paraffin embedded (human 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 (phospho-SMAD5 (Ser463+Ser465)) Monoclonal Antibody, Unconjugated (TMAB-11304) at 1: 200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
2. 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 (phospho-SMAD5 (Ser463+Ser465)) Monoclonal Antibody, Unconjugated (TMAB-11304) at 1: 200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
3. Paraformaldehyde-fixed, paraffin embedded (rat 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 (phospho-SMAD5 (Ser463+Ser465)) Monoclonal Antibody, Unconjugated (TMAB-11304) at 1: 200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
4. Paraformaldehyde-fixed, paraffin embedded (human breast carcinoma); 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 (phospho-SMAD5 (Ser463+Ser465)) Monoclonal Antibody, Unconjugated (TMAB-11304) at 1: 200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
5. Paraformaldehyde-fixed, paraffin embedded (mouse colon); 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 (phospho-SMAD5 (Ser463+Ser465)) Monoclonal Antibody, Unconjugated (TMAB-11304) at 1: 200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
6. Paraformaldehyde-fixed, paraffin embedded (mouse 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 (phospho-SMAD5 (Ser463+Ser465)) Monoclonal Antibody, Unconjugated (TMAB-11304) at 1: 200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
7. Paraformaldehyde-fixed, paraffin embedded (Human skin cancer); 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 (phospho-SMAD5 (Ser463+Ser465)) Monoclonal Antibody, Unconjugated (TMAB-11304) at 1: 200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
8. Paraformaldehyde-fixed, paraffin embedded (human smooth muscle); 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 (phospho-SMAD5 (Ser463+Ser465)) Monoclonal Antibody, Unconjugated (TMAB-11304) at 1: 200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
9. Paraformaldehyde-fixed, paraffin embedded (rat colon); 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 (phospho-SMAD5 (Ser463+Ser465)) Monoclonal Antibody, Unconjugated (TMAB-11304) at 1: 200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
10. 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 (phospho-SMAD5 (Ser463+Ser465)) Monoclonal Antibody, Unconjugated (TMAB-11304) at 1: 200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
11. Paraformaldehyde-fixed, paraffin embedded (human tonsil); 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 (phospho-SMAD5 (Ser463+Ser465)) Monoclonal Antibody, Unconjugated (TMAB-11304) at 1: 200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
12. Paraformaldehyde-fixed, paraffin embedded Mouse Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-SMAD5 (Ser463+Ser465) Monoclonal Antibody, Unconjugated (TMAB-11304) at 1: 200 overnight at 4°C, followed by conjugation to the Goat Anti-Rabbit IgG H&L Secondary Antibody-HRP and DAB staining.
13. Paraformaldehyde-fixed, paraffin embedded Rat Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-SMAD5 (Ser463+Ser465) Monoclonal Antibody, Unconjugated (TMAB-11304) at 1: 200 overnight at 4°C, followed by conjugation to the Goat Anti-Rabbit IgG H&L Secondary Antibody-HRP and DAB staining.
14. Paraformaldehyde-fixed, paraffin embedded Human Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-SMAD5 (Ser463+Ser465) Monoclonal Antibody, Unconjugated (TMAB-11304) at 1: 200 overnight at 4°C, followed by conjugation to the Goat Anti-Rabbit IgG H&L Secondary Antibody-HRP and DAB staining.
15. Paraformaldehyde-fixed, paraffin embedded Mouse Placenta; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-SMAD5 (Ser463+Ser465) Monoclonal Antibody, Unconjugated (TMAB-11304) at 1: 200 overnight at 4°C, followed by conjugation to the Goat Anti-Rabbit IgG H&L Secondary Antibody-HRP and DAB staining.
16. Paraformaldehyde-fixed, paraffin embedded Human Placenta; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-SMAD5 (Ser463+Ser465) Monoclonal Antibody, Unconjugated (TMAB-11304) at 1: 200 overnight at 4°C, followed by conjugation to the Goat Anti-Rabbit IgG H&L Secondary Antibody-HRP and DAB staining.
17. Paraformaldehyde-fixed, paraffin embedded Mouse Heart; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-SMAD5 (Ser463+Ser465) Monoclonal Antibody, Unconjugated (TMAB-11304) at 1: 200 overnight at 4°C, followed by conjugation to the Goat Anti-Rabbit IgG H&L Secondary Antibody-HRP and DAB staining.
18. Paraformaldehyde-fixed, paraffin embedded Rat Heart; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-SMAD5 (Ser463+Ser465) Monoclonal Antibody, Unconjugated (TMAB-11304) at 1: 200 overnight at 4°C, followed by conjugation to the Goat Anti-Rabbit IgG H&L Secondary Antibody-HRP and DAB staining.
19. Paraformaldehyde-fixed, paraffin embedded Human Heart; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-SMAD5 (Ser463+Ser465) Monoclonal Antibody, Unconjugated (TMAB-11304) at 1: 200 overnight at 4°C, followed by conjugation to the Goat Anti-Rabbit IgG H&L Secondary Antibody-HRP and DAB staining.
20. Paraformaldehyde-fixed, paraffin embedded Mouse Skin; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-SMAD5 (Ser463+Ser465) Monoclonal Antibody, Unconjugated (TMAB-11304) at 1: 200 overnight at 4°C, followed by conjugation to the Goat Anti-Rabbit IgG H&L Secondary Antibody-HRP and DAB staining.
21. Paraformaldehyde-fixed, paraffin embedded Rat Skin; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-SMAD5 (Ser463+Ser465) Monoclonal Antibody, Unconjugated (TMAB-11304) at 1: 200 overnight at 4°C, followed by conjugation to the Goat Anti-Rabbit IgG H&L Secondary Antibody-HRP and DAB staining.
22. Paraformaldehyde-fixed, paraffin embedded Human Skin; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-SMAD5 (Ser463+Ser465) Monoclonal Antibody, Unconjugated (TMAB-11304) at 1: 200 overnight at 4°C, followed by conjugation to the Goat Anti-Rabbit IgG H&L Secondary Antibody-HRP and DAB staining.
23. 25 μg total protein per lane of various lysates (see on figure) probed with phospho-SMAD5 (Ser463+Ser465) monoclonal antibody, unconjugated (TMAB-11304) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r. T. for 60 min.

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; ICC/IF: 1:50-200; IF: 1:100-500

Monoclonal

KLH conjugated synthesised phosphopeptide: human SMAD5 around the phosphorylation site of Ser463 + Ser465

Human

SMAD5

Mothers against decapentaplegic homolog 5

Transcriptional modulator activated by BMP (bone morphogenetic proteins) type 1 receptor kinase. SMAD5 is a receptor-regulated SMAD (R-SMAD).