VALIDATION IMAGES
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 Doublecortin (Ser47) Polyclonal Antibody, Unconjugated (bs-3113R) at 1:400 overnight at 4°C, DAB staining.
U251 cell lysates probed with Doublecortin (Ser47) Polyclonal Antibody, Unconjugated (bs-3113R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
Human HepG2 Cells were fixed with 2% PFA (10min at room temperature) and then permeabilized with 0.2% PBST for 30min at room temperature. The cells were then incubated in 2% BSA + 10% goat serum to block non-specific protein-protein interactions for 15 min at room temperature. Cells were stained with Doublecortin (Ser47) Polyclonal Antibody, Unconjugated (bs-3113R) at 1:100 for 30 min at room temperature. The secondary antibody, Goat anti-rabbit IgG-FITC (bs-0295G-FITC), was used for 40 min at room temperature. Primary Antibody staining (green) is compared to unstained cells (black), secondary only (blue) and isotype control (bs-0295P; orange).
Lane 1: Mouse Cerebrum lysates; Lane 2: Rat Cerebrum lysates probed with Doublecortin (Ser47) Polyclonal Antibody, Unconjugated (bs-3113R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
SHSY5Y cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (Phospho-Doublecortin (Ser128)) polyclonal Antibody, Unconjugated (bs-3113R) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.