Formalin-fixed and paraffin-embedded human hepatocarcinoma tissue reacted with hUVRAG , 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.
Immunofluorescence staining of Autophagy UVRAG antibody on Methanol-fixed HeLa cells. Data courtesy of Dr. Eeva-Liisa Eskelinen, University of Helsinki,Finland.
Fluorescent image of U251 cells stained with UVRAG 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 UVRAG 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, 1h). Nuclei were counterstained with Hoechst 33342 (blue) (10 ug/ml, 5 min). UVRAG immunoreactivity is localized to autophagic vacuoles in the cytoplasm of U251 cells.
Immunoprecipitation and western blot of anti-UVRAG antibody in 293T cells. UVRAG is immunoprecipitated (Lane 2) and detected in 293T cell transiently transfected with mouse UVRAG (Lane 1). Detection of endogenous UVRAG is shown in 293T cells (Lane 3) but is reduced by UVRAG siRNA transfection (Lane 4). Data courtesy of Dr. Hong-Gang Wang, Moffitt Cancer Center, Tampa, FL.