bsm-52206R [Primary Antibody]
phospho-SMAD5 (Ser463 + Ser465) Recombinant Antibody
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Host: Rabbit

Target Protein: phospho-SMAD5 (Ser463 + Ser465)

Modification Site: S463/465

Clonality: Recombinant

Isotype: IgG

Entrez Gene: 4090

Swiss Prot: Q99717

Source: KLH conjugated synthesised phosphopeptide derived from human SMAD5 around the phosphorylation site of Ser463 + Ser465:IS(p-S)V(p-S).

Purification: Purified by Protein A.

Storage Buffer: 0.01M TBS (pH 7.4), 1% BSA, 0.02% Proclin 300, and 50% Glycerol

Storage: Shipped at 4C. Store at -20C for one year. Avoid repeated freeze/thaw cycles.

Background:

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

Size: 100µL

Concentration: Lot Dependent

Cross Reactive Species: Human
Mouse
Rat

For research use only. Not intended for diagnostic or therapeutic use.

VALIDATION IMAGES

Paraformaldehyde-fixed, paraffin embedded Mouse liver; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with Smad5(S463/465) (1B8) Monoclonal Antibody, Unconjugated (bsm-52206R) at 1:200 overnight at 4°C, DAB staining.


Paraformaldehyde-fixed, paraffin embedded Mouse colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with Smad5(S463/465) (1B8) Monoclonal Antibody, Unconjugated (bsm-52206R) at 1:200 overnight at 4°C, DAB staining.


Paraformaldehyde-fixed, paraffin embedded Mouse brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with Smad5(S463/465) (1B8) Monoclonal Antibody, Unconjugated (bsm-52206R) at 1:200 overnight at 4°C, DAB staining.


Paraformaldehyde-fixed, paraffin embedded Rat liver; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with Smad5(S463/465) (1B8) Monoclonal Antibody, Unconjugated (bsm-52206R) at 1:200 overnight at 4°C, DAB staining.


Paraformaldehyde-fixed, paraffin embedded Rat colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with Smad5(S463/465) (1B8) Monoclonal Antibody, Unconjugated (bsm-52206R) at 1:200 overnight at 4°C, DAB staining.


Paraformaldehyde-fixed, paraffin embedded Rat brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with Smad5(S463/465) (1B8) Monoclonal Antibody, Unconjugated (bsm-52206R) at 1:200 overnight at 4°C, DAB staining.


Paraformaldehyde-fixed, paraffin embedded Human skin cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with Smad5(S463/465) (1B8) Monoclonal Antibody, Unconjugated (bsm-52206R) at 1:200 overnight at 4°C, DAB staining.


Paraformaldehyde-fixed, paraffin embedded Human tonsil; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with Smad5(S463/465) (1B8) Monoclonal Antibody, Unconjugated (bsm-52206R) at 1:200 overnight at 4°C, DAB staining.


Paraformaldehyde-fixed, paraffin embedded Human brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with Smad5(S463/465) (1B8) Monoclonal Antibody, Unconjugated (bsm-52206R) at 1:200 overnight at 4°C, DAB staining.


Paraformaldehyde-fixed, paraffin embedded Human smooth muscle; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with Smad5(S463/465) (1B8) Monoclonal Antibody, Unconjugated (bsm-52206R) at 1:200 overnight at 4°C, DAB staining.


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 (bsm-52206R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.


Paraformaldehyde-fixed, paraffin embedded Human breast cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with Smad5(S463/465) (1B8) Monoclonal Antibody, Unconjugated (bsm-52206R) at 1:200 overnight at 4°C, DAB staining.


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 (bsm-52206R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.


Paraformaldehyde-fixed, paraffin embedded Mouse brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with Smad5(S463/465) (1B8) Monoclonal Antibody, Unconjugated (bsm-52206R) at 1:200 overnight at 4°C, DAB staining.


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 (bsm-52206R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.


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 (bsm-52206R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.


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 (bsm-52206R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.


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 (bsm-52206R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.


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 (bsm-52206R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.


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 (bsm-52206R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.


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 (bsm-52206R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.


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 (bsm-52206R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.


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 (bsm-52206R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.


25 ug total protein per lane of various lysates (see on figure) probed with phospho-SMAD5 (Ser463 + Ser465) monoclonal antibody, unconjugated (bsm-52206R) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.