Minute™ Cytosolic Proteasome Enrichment Kit (20 preps)

Manual & Protocol | MSDS

The proteasome is a non-lysosomal protein degradation system that requires metabolic energy and ubiquitin polymerization. The 26S proteasome is made up of two subcomplexes (a 20S proteasome and one or two19S regulatory particles). Rapid and gentle isolation of proteasomes from cells/tissues is essential for the study of proteasome structure and function. Proteasomes are mainly localized in the cytosol but are also detected in the nucleus. Traditionally, ultracentrifugation and affinity isolation are the most common methods for isolation of proteasomes. These methods, though relatively effective, are usually time-consuming and low-yielding. Many affinity-based methods require harsh elution conditions that may affect isolated proteasome activity 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 proteasomes from cytosol by first removing nuclei and the majority of organelles, followed by preferential precipitation of cytosolic proteasomes. Nuclear proteasomes are excluded with this kit (Nuclear Proteasome Enrichment Kit is available under Cat # PN-041). The protocol is simple, rapid, and high-yielding. The gentle protocol maintains the association of proteasomes with ubiquitin and other proteins. This kit is useful for the study of proteasome structure and function. The enriched proteasomes can also be used as a starting material for affinity purifications.

Kit Components:

 References (2)

1.  Zhang, C. Y., Zhang, R., Zhang, L., Wang, Z. M., Sun, H. Z., Cui, Z. G., & Zheng, H. C. (2023). Regenerating gene 4 promotes chemoresistance of colorectal cancer by affecting lipid droplet synthesis and assembly. World Journal of Gastroenterology, 29(35), 5104-5124.

2.  Rao, N. R., Upadhyay, A., & Savas, J. N. (2024). Derailed protein turnover in the aging mammalian brain. Molecular Systems Biology, 1-20.