All lanes : Anti-LAMP1 antibody - Lysosome Marker (ab79821) at 1/2000 dilutionLane 1 : Normal Rat Cortical LysateLane 2 : Normal Human Cortical LysateLane 3 : Parkinson's disease dementia Cortical LysateLane 4 : Dementia with Lewy Bodies Cortical LysateLane 5 : Alzheimer's Disease Cortical LysatesLysates/proteins at 20 µg per lane.SecondaryAnti Rabbit IgG conjugated to Alexa Fluor at 1/5000 dilutionPerformed under reducing conditions.
ICC/IF image of ab79821 stained MCF7 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab79821, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% formaldehyde fixed (10 min) HeLa cells at 5µg/ml.