Tissue: Human colon cancer Section type: Formalin-fixed & Paraffinembedded section Retrieval method: High temperature and high pressure Retrieval buffer: Tris/EDTA buffer, pH 9.0 Primary Ab dilution: 1:500 Primary Ab incubation condition: 1 hour at room temperature Secondary Ab: Anti-Rabbit and Mouse Polymer HRP (Ready to use) Counter stain: Hematoxylin (Blue) Comment: Color brown is the positive signal for bsm-60709R


Tissue: Human lung squamous carcinoma Section type: Formalin-fixed & Paraffinembedded section Retrieval method: High temperature and high pressure Retrieval buffer: Tris/EDTA buffer, pH 9.0 Primary Ab dilution: 1:500 Primary Ab incubation condition: 1 hour at room temperature Secondary Ab: Anti-Rabbit and Mouse Polymer HRP (Ready to use) Counter stain: Hematoxylin (Blue) Comment: Color brown is the positive signal for bsm-60709R


Blocking buffer: 5% NFDM/TBST Primary Ab dilution: 1:10000 Primary Ab incubation condition: 2 hours at room temperature Secondary Ab: Goat Anti-Rabbit IgG H&L (HRP) Lysate: (-): HCT-116, (+): HCT-116 +chloroquine (50 mM, 12h) Protein loading quantity: 20 μg Exposure time: 30 s Predicted MW: 48 kDa Observed MW: 62 kDa


Paraformaldehyde-fixed, paraffin embedded (mouse spleen); 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 (SQSTM1/p62) Monoclonal Antibody, Unconjugated (bsm-60709R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.


Blocking buffer: 5% NFDM/TBST Primary Ab dilution: 1:10000 Primary Ab incubation condition: 2 hours at room temperature Secondary Ab: Goat Anti-Rabbit IgG H&L (HRP) Lysate: (-): HeLa, (+): HeLa+chloroquine (50 mM, 12h) Protein loading quantity: 20 μg Exposure time: 30 s Predicted MW: 48 kDa Observed MW: 62 kDa


Paraformaldehyde-fixed, paraffin embedded (rat spleen); 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 (SQSTM1/p62) Monoclonal Antibody, Unconjugated (bsm-60709R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.


Paraformaldehyde-fixed, paraffin embedded (human spleen); 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 (SQSTM1/p62) Monoclonal Antibody, Unconjugated (bsm-60709R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.


Paraformaldehyde-fixed, paraffin embedded (human stomach); 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 (SQSTM1/p62) Monoclonal Antibody, Unconjugated (bsm-60709R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.


Paraformaldehyde-fixed, paraffin embedded (human liver carcinoma); 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 (SQSTM1/p62) Monoclonal Antibody, Unconjugated (bsm-60709R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.


25 ug total protein per lane of various lysates (see on figure) probed with SQSTM1/p62 monoclonal antibody, unconjugated (bsm-60709R) at 1:5000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.