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enzymes contained in lysosomes play a vital role in digesting excess or worn-out organelles, food particles

and engulfed viruses or bacteria. Lysosomes are relatively large organelles ranging in size from 0.1 to 1.2

um. The ability to isolate lysosomes is an important first step in studies of autophagy, protein degradation

and protein recycling in a cell. Traditional methods for isolatinglysosomes are based on density-gradient

ultracentrifugation. A large amount of starting material is required and the methods are tedious and time-

consuming and contain significant cross-contamination. Currently, all commercial kits for lysosome isolation

are based on methods developed in the 1970s. Unlike any other lysosome isolation kit on the market, our kit

employs a patented spin-column-based technology that is simple, rapid and efficient. The amount of starting

cells/tissues required is much smaller thanthat of traditional methods. This kit can significantly enrich

lysosomes from cultured cells or tissues without using a Dounce homogenizer and ultracentrifugation. The

whole protocol can be done in less than 1.5 hours.

Isolation of lysosome fraction from frozen mouse liver and kidney.  A. Ponseau red staining of transferred

Western blot. B. Western blot with different marker antibodies. Anti-LIMPII/Ipg85 for lysosome, anti-ER and

anti-Golgi antibodies were purchased from NovusBio. Anti-mitochondria and nuclear marker antibodies were

from Abcam and Boster, respectively. T, Total tissue lysate, L, Lysosome fraction.

Isolation of lysosome fraction from fresh mouse tissues.  A. Ponseau S stained blot. B. Western blot with

anit-lysosome marker antibodies(Novus, NB400-129SS).  Lane 1. Liver, 2. Kidney, 3. Spleen, 4. Lung