Application
Fluorescent image of U251 cells stained with cleaved LC3B antibody. U251 cells were treated with Chloroquine (50 μM,16h), then fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.2%, 30 min). Cells were then incubated with AP1806a cleaved LC3B 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 μg/ml, 5 min). LC3 immunoreactivity is localized to autophagic vacuoles in the cytoplasm of U251 cells.
Application
Time course study of mouse leukaemic monocyte macrophage cells treated with U18666A, a drug that causes cholesterol and lipid storage in cells, thereby blocking fusion between late endosomes and lysosomes. Cleaved-LC3 (APG8b) antibody (Cat AP1806a) detected punctuate staining indicative of autophagic vacuole or phagosome structures. Data courtesy of Dr. Barry Boland, Department of Pharmacology, Oxford University.