| Western blot (WB): | 1:500-2000 |
| Flow Cytometry (Fixed): | 1:50-200 |
| Enzyme linked immunosorbent assay (ELISA): | 1:100-1000 |

Western blot analysis of ATG4B using anti-ATG4B antibody (A02885-1). The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: Hela whole cell lysates,
Lane 2: 293T whole cell lysates,
Lane 3: HepG2 whole cell lysates,
Lane 4: Jurkat whole cell lysates,
Lane 5: RT4 whole cell lysates,
Lane 6: MCF-7 whole cell lysates,
Lane 7: MOLF-4 whole cell lysates,
Lane 8: HEL whole cell lysates,
Lane 9: rat brain tissue lysates,
Lane 10: rat liver tissue lysates,
Lane 11: RH35 whole cell lysates.
After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-ATG4B antigen affinity purified polyclonal antibody (A02885-1) at a dilution of 1:1000 and probed with a goat anti-rabbit IgG-HRP secondary antibody (Catalog # BA1054). The signal is developed using ECL Plus Western Blotting Substrate (Catalog # AR1197). A specific band was detected for ATG4B at approximately 48 kDa. The expected band size for ATG4B is at 44 kDa.

Flow Cytometry analysis of Jurkat cells using anti-ATG4B antibody (A02885-1).
Overlay histogram showing Jurkat cells stained with A02885-1 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ATG4B Antibody (A02885-1) at 1:100 dilution for 30 min at 20°C. Fluoro488 conjugated goat anti-rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG at 1:100 dilution used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Western blot analysis of ATG4B using anti-ATG4B antibody (A02885-1). The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: Hela whole cell lysates,
Lane 2: 293T whole cell lysates,
Lane 3: HepG2 whole cell lysates,
Lane 4: Jurkat whole cell lysates,
Lane 5: RT4 whole cell lysates,
Lane 6: MCF-7 whole cell lysates,
Lane 7: MOLF-4 whole cell lysates,
Lane 8: HEL whole cell lysates,
Lane 9: rat brain tissue lysates,
Lane 10: rat liver tissue lysates,
Lane 11: RH35 whole cell lysates.
After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-ATG4B antigen affinity purified polyclonal antibody (A02885-1) at a dilution of 1:1000 and probed with a goat anti-rabbit IgG-HRP secondary antibody (Catalog # BA1054). The signal is developed using ECL Plus Western Blotting Substrate (Catalog # AR1197). A specific band was detected for ATG4B at approximately 48 kDa. The expected band size for ATG4B is at 44 kDa.

Flow Cytometry analysis of Jurkat cells using anti-ATG4B antibody (A02885-1).
Overlay histogram showing Jurkat cells stained with A02885-1 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ATG4B Antibody (A02885-1) at 1:100 dilution for 30 min at 20°C. Fluoro488 conjugated goat anti-rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG at 1:100 dilution used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.



