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The proteasome is a non-lysosome protein degradation system that requires metabolic energy and

polymerization of ubiquitin. The 26S proteasome is made up of two subcomplexes (a 20S proteasome and

one or two19S regulatory particles). Rapid and gentle isolation of proteasome from cells/tissues is essential

for the studies of proteasome structure and function. The proteasomes have been found mainly localized in

cytosol but also detected in nucleus. Traditionally, ultracentrifugation and affinityisolation are the most

common methods for isolation of proteasomes. These methods, though relatively effective, are usually time

consuming with low yield. Many affinity-based methods require harsh elution condition that may affect the

activity of isolated proteasomes and limit certain downstream applications. To overcome these

shortcomings, we have developed a spin-column based technology using a proprietary precipitation buffer

for enrichment of proteasome from cytosol by first removing nuclei and majority oInventBiotechnologies Inc.f

organelles followed by preferential precipitation of cytosolic proteasomes. The nuclear proteasomes are

excluded with this kit. The protocol is simple and rapid with high yield. The gentle protocol maintains the

association of proteasomes with ubiquitin and other proteins. This kit is useful for the studies of proteasome

structure and function. The enriched proteasomes can also be used as a starting material for affinity

purifications.