Sample:
Lane 1: Mouse Cerebrum Lysates
Lane 2: Mouse Cerebellum Lysates
Lane 3: Rat Cerebrum Lysates
Lane 4: Rat Cerebellum Lysates
Primary: Anti-Synaptophysin (bsm-52379R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 34 kDa
Observed band size: 34 kDa

Paraformaldehyde-fixed, paraffin embedded (mouse pancreas); 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 (Synaptophysin) Monoclonal Antibody, Unconjugated (bsm-52379R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (Human left parietal lobe); 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; Incubation with (Synaptophysin) Monoclonal Antibody, Unconjugated (bsm-52379R) at 1:100 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023)instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (human pancreas); 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; Incubation with (Synaptophysin) Monoclonal Antibody, Unconjugated (bsm-52379R) at 1:100 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023)instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (rat cerebellum); 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; Incubation with (Synaptophysin) Monoclonal Antibody, Unconjugated (bsm-52379R) at 1:100 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023)instructionsand 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; Incubation with (Synaptophysin) Monoclonal Antibody, Unconjugated (bsm-52379R) at 1:100 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023)instructionsand 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; Incubation with (Synaptophysin) Monoclonal Antibody, Unconjugated (bsm-52379R) at 1:100 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023)instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (mouse cerebellum); 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; Incubation with (Synaptophysin) Monoclonal Antibody, Unconjugated (bsm-52379R) at 1:100 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023)instructionsand 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; Incubation with (Synaptophysin) Monoclonal Antibody, Unconjugated (bsm-52379R) at 1:100 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023)instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (human cerebellum); 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; Incubation with (Synaptophysin) Monoclonal Antibody, Unconjugated (bsm-52379R) at 1:100 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023)instructionsand 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; Incubation with (Synaptophysin) Monoclonal Antibody, Unconjugated (bsm-52379R) at 1:100 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023)instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (rat pancreas); 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 (Synaptophysin) Monoclonal Antibody, Unconjugated (bsm-52379R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand 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 (Synaptophysin) Monoclonal Antibody, Unconjugated (bsm-52379R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

PC12 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 (Synaptophysin) Monoclonal Antibody, Unconjugated (bsm-52379R) 1:50, 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 nuclei.

Blank control:N2A.
Primary Antibody (green line): Rabbit Anti-Synaptophysin antibody (bsm-52379R)
Dilution: 1:50;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-AF488
Dilution: 1:1000.
Protocol
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. Acquisition of 20,000 events was performed.