VALIDATION IMAGES
U2-OS 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 (SQSTM1/p62) polyclonal Antibody, Unconjugated (bs-55207R) 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.
Paraformaldehyde-fixed, paraffin embedded Rat ovarian; 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 SQSTM1/p62 Polyclonal Antibody, Unconjugated (bs-55207R) at 1:100 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Human appendix; 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 SQSTM1/p62 Polyclonal Antibody, Unconjugated (bs-55207R) at 1:100 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Mouse liver; 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 SQSTM1/p62 Polyclonal Antibody, Unconjugated (bs-55207R) at 1:100 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Human esophagus; 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 SQSTM1/p62 Polyclonal Antibody, Unconjugated (bs-55207R) at 1:100 overnight at 4°C, DAB staining.
Lane 1: 293T cell lysates; Lane 2: SQSTM1/p62 knockout (KO) 293T cell lysates probed with SQSTM1/p62 Polyclonal Antibody, Unconjugated (bs-55207R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
Lane 1: 293T cell lysates; Lane 2: Hela cell lysates; Lane 3: U87-MG cell lysates probed with SQSTM1/p62 Polyclonal Antibody, Unconjugated (bs-55207R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.