Formalin-fixed and paraffin-embedded human cancer tissue reacted with the primary antibody, which was peroxidase-conjugated to the secondary antibody, followed by DAB staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated. BC = breast carcinoma; HC = hepatocarcinoma.
5Y cells were pretreated with 5nM bafilomycin for 24hr and fixed in 4% of paraformaldehyde. Treatment with antibody at dilution 1:100. Data courtesy of Jianhui Zhu, MD, PhD & Charleen T. Chu, MD, PhD, University of Pittsburgh School of Medicine.
Fluorescent image of U251 cells stained with cleaved LC3A antibody. U251 cells were fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.2%, 30 min). Cells were then incubated with cleaved LC3A primary antibody (1:100, 2 h at room temperature). For secondary antibody, Alexa Fluor 488 conjugated donkey anti-rabbit antibody (green) was used (1:1000, 1h). Nuclei were counterstained with Hoechst 33342 (blue) (10 ug/ml, 5 min). LC3 immunoreactivity is localized to autophagic vacuoles in the cytoplasm of U251 cells.
Fluorescent image of U251 cells stained with cleaved LC3A antibody. U251 cells were treated with Chloroquine (50 mu M,16h), then fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.2%, 30 min). Cells were then incubated cleaved LC3A primary antibody (1:100, 2 h at room temperature). For secondary antibody, Alexa Fluor 488 conjugated donkey anti-rabbit antibody (green) was used (1:1000, 1h). Nuclei were counterstained with Hoechst 33342 (blue) (10 ug/ml, 5 min). LC3 immunoreactivity is localized to autophagic vacuoles in the cytoplasm of U251 cells.
Western blot of anti-cleaved-LC3 (APG8a) antibody in mouse brain tissue lysate. Cleaved-LC3 (APG8a) was detected using the purified antibody.