Western blot analysis of SQSTM1 / p62 on different lysates with Rabbit anti-SQSTM1 / p62 antibody (HA721171) at 1/5;000 dilution. Lane 1: SH-SY5Y cell lysate Lane 2: PANC-1 cell lysate Lane 3: HeLa cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 48 kDa Observed band size: 62 kDa Exposure time: 1 minute; ECL: K1801; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721171) at 1/5;000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100;000 dilution was used for 1 hour at room temperature.
Western blot analysis of SQSTM1 / p62 on different lysates with Rabbit anti-SQSTM1 / p62 antibody (HA721171) at 1/2;000 dilution. Lane 1: A549-si NT cell lysate Lane 2: A549-si SQSTM1 / p62 cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 48 kDa Observed band size: 62 kDa Exposure time: 17 seconds; ECL: K1801; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721171) at 1/2;000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50;000 dilution was used for 1 hour at room temperature.
Immunocytochemistry analysis of C2C12 cells labeling SQSTM1 / p62 with Rabbit anti-SQSTM1 / p62 antibody (HA721171) at 1/1;000 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature; permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature; then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-SQSTM1 / p62 antibody (HA721171) at 1/1;000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488; HA1121) was used as the secondary antibody at 1/1;000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2; red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594; HA1126) was used as the secondary antibody at 1/1;000 dilution.
Immunocytochemistry analysis of PC-12 cells labeling SQSTM1 / p62 with Rabbit anti-SQSTM1 / p62 antibody (HA721171) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature; permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature; then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-SQSTM1 / p62 antibody (HA721171) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488; HA1121) was used as the secondary antibody at 1/1;000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2; red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594; HA1126) was used as the secondary antibody at 1/1;000 dilution.
