VALIDATION IMAGES
Paraformaldehyde-fixed, paraffin embedded Mouse Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with LC3A Monoclonal Antibody, Unconjugated(bsm-62899R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023)and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Rat Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with LC3A Monoclonal Antibody, Unconjugated(bsm-62899R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023)and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with LC3A Monoclonal Antibody, Unconjugated(bsm-62899R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023)and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Rat Cerebellum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with LC3A Monoclonal Antibody, Unconjugated(bsm-62899R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023)and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Cerebellum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with LC3A Monoclonal Antibody, Unconjugated(bsm-62899R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023)and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Glioma; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with LC3A Monoclonal Antibody, Unconjugated(bsm-62899R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023)and DAB (C-0010) staining.
4% Paraformaldehyde-fixed Hela (treated with chloroquine (50uM,overnight) ) (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (LC3A) monoclonal Antibody, unconjugated (bsm-62899R) 1:100, 90 min at 37°C; followed by conjugated Goat Anti-Rabbit IgG antibody (green, bs-60295G-BF488) at 37°C for 90 min, DAPI (blue, C02-04002) was used to stain the cell nuclei. PBS instead of the primary antibody was used as the blank control.
25 ug total protein per lane of various lysates (see on figure) probed with LC3A monoclonal antibody, unconjugated (bsm-62899R) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.