ATG7 Antibody

Staining APG7L in human liver tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 3% BSA for 0.5 hour at room temperature; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody (1/25) for 1 hours at 37°C. A undiluted biotinylated goat polyvalent antibody was used as the secondary antibody.

Fluorescent image of U251 cells stained with ATG7 antibody. U251 cells were treated with Chloroquine (50 μM, 16 h), then fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.2%, 30 min). Cells were then incubated with ATG7 primary antibody (1:200, 2 h at room temperature). For secondary antibody, Alexa Fluor 488 conjugated donkey anti-rabbit antibody (green) was used (1:1000, 1 h). Nuclei were counterstained with Hoechst 33342 (blue) (10 μg/ml, 5 min). ATG7 immunoreactivity is localized to autophagic vacuoles in the cytoplasm of U251 cells.

Western blot analysis of anti-hAPG7L-R509 Pab in 293 cell line lysates transiently transfected with the ATG7 gene (2 ug/lane). hAPG7L-R509 (arrow) was detected using the purified Pab.

All lanes: Anti-APG7L Antibody (R509) at 1:1000 dilution. Lane 1: Ba/F3 whole cell lysate. Lane 2: Caki-1 whole cell lysate. Lane 3: HepG2 whole cell lysate. Lysates/proteins at 20 µg per lane. Secondary Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/10000 dilution. Predicted band size: 78 kDa. Blocking/Dilution buffer: 5% NFDM/TBST.