Formalin-fixed and paraffin-embedded human brain tissue reacted with AIFM1 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.
Fluorescent image of U251 cells stained with AIFM1 antibody. U251 cells were fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.2%, 30 min). Cells were then incubated AIFM1 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). Cytoplasmic actin was counterstained with Alexa Fluor 555 (red) conjugated Phalloidin (5.25 mu M, 25 min). Pictures were taken on a Biorevo microscope (BZ-900, Keyence). Nuclei were counterstained with Hoechst 33342 (blue) (10 ug/ml, 5 min). AIFM1 immunoreactivity is localized to the cytoplasm of U251 cells.
Fluorescent confocal image of U251 cells stained with AIFM1 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 AIFM1 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). AIFM1 immunoreactivity is localized to the cytoplasm of U251 cells.
Western blot of AIFM1 Antibody in 293 cell line lysates (35 ug/lane). AIFM1 (arrow) was detected using the purified antibody.
AIFM1 Antibody flow cytometry of 293 cells (right histogram) compared to a negative control cell (left histogram). FITC-conjugated goat-anti-rabbit secondary antibodies were used for the analysis.