Santa Cruz Biotechnology
Nile Red
sc-203747
Nile Red (NR) is a lipophilic dye used to stain lipid droplets in cells, and also protein aggregates. I used NR to visualize aggresomes in chloroquine-treated MDA231 breast adenocarcinoma cells.
Immunofluoresence
MDA231 breast adenocarcinoma cells
Stock solution of Nile Red: 10 mg/ml in acetone or water-free DMSO. Dilute 1:1000 in PBS for staining fixed, permabilized cells, for 15 mins using 0.5 ml of diluted dye solution. This was followed by 3x five minute washes in PBS. Imaging was captured on a Nikon Eclipse Ti inverted microscope using NIS Elements software.
Store stock solution of NR at RT in a light protected bottle.
As can be seen in Fig 1, cells treated overnight in 25 uM chloroquine (an aggresome inducer) display distinct cytoplasmic inclusions typical of aggresomes. Fluorescence is seen in the TRITC/PE channel.
N/A
Easy to use, in expensive, very bright staining
Fluoresence extends into FITC channels, making two color immunofluoresence problematic. DAPI channels were unaffected.
An excellent and inexpensive dye.