Immunocytochemistry/Immunofluorescence analysis of BACE2 (green) showing staining in the nucleus and cytoplasm of A549 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were incubated with ab5670 in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4 ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a flourescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.
All lanes : Anti-BACE2 antibody (ab5670) at 1/500 dilutionLane 1 : Mouse spleen cell lysateLane 2 : SH-SY5Y cell lysateLane 3 : HL60 cell lysateLane 4 : HeLa cell lysateLysates/proteins at 25 µg per lane.