25 ug total protein per lane of various lysates (see on figure) probed with Synaptophysin monoclonal antibody, unconjugated (bsm-52379R) at 1:2000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.

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 Human Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Synaptophysin Monoclonal Antibody, Unconjugated(bsm-52379R) 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 Pancreas; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Synaptophysin Monoclonal Antibody, Unconjugated(bsm-52379R) 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 Pancreas; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Synaptophysin Monoclonal Antibody, Unconjugated(bsm-52379R) 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 Pancreas; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Synaptophysin Monoclonal Antibody, Unconjugated(bsm-52379R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) 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.