VALIDATION IMAGES
HeLa cells were stained with alpha-Synuclein Polyclonal Antibody, Unconjugated(bs-0968R) at 1:500 in PBS and incubated for two hours at 37°C followed by Goat Anti-Rabbit IgG (H+L) Cy3 conjugated secondary antibody. DAPI staining of the nucleus was done and then detected.
Paraformaldehyde-fixed, paraffin embedded Rat spinal cord; 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 Alpha-Synuclein Polyclonal Antibody, Unconjugated (bs-0968R) at 1:200 overnight at 4°C, DAB staining.
25 ug total protein per lane of various lysates (see on figure) probed with Alpha-Synuclein polyclonal antibody, unconjugated (bs-0968R) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.
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 Alpha-Synuclein Polyclonal Antibody, Unconjugated (bs-0968R) 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 Alpha-Synuclein Polyclonal Antibody, Unconjugated (bs-0968R) 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 Alpha-Synuclein Polyclonal Antibody, Unconjugated (bs-0968R) at 1:200 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Mouse spinal cord; 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 Alpha-Synuclein Polyclonal Antibody, Unconjugated (bs-0968R) at 1:200 overnight at 4°C, DAB staining.