The nuclear envelope is a very complex membrane-protein system that is notoriously difficult to isolate and purify because of its connection to nuclear and cytoplasmic components. Traditional methods of nuclear envelope isolation and purification require a large amount of starting material and a long, tedious procedure. Minute™ Nuclear Envelope Protein Extraction Kit is the first commercial kit designed to rapidly isolate nuclear envelope and its associated proteins in native form without using density-gradient and ultracentrifugation. Due to the use of protein extraction filter cartridges, the nuclear envelope protein isolation is simple and user-friendly. The nuclear envelope proteins are significantly enriched in the final prep. Unlike traditional methods that require large amounts of starting cells/tissues, this kit starts with only 10-20 million cells, and the buffers are detergent and EDTA free. The procedure can be completed in less than 45 minutes with a final yield of 10-50 µg protein/sample.
A. SDS-PAGE profiles and Western blottings of isolated mouse spleen cells. Lanes 1 and 2, total cell lysate; Lane 3, nuclear envelope.
B. SDS-PAGE profiles and Western blottings of Rate (REL-6TN) and human (MCF-7) cell lines. Lane 1, REL-6TN total cell lysate; Lane 2, REL-6TN nuclear envelope. Lane 3, MCF-7 total cell lysate; Lane 4, MCF-7 nuclear envelope. Nuclear envelope marker Lamin B1 (ab 16048, ABcam, Cambridge MA) and GAPDH (G9545, Sigma) were visualized by a color metric substrate (Bio-Rad).
|Buffer A||25 ml|
|Buffer B||25 ml|
|Filter Cartridges||50 Units|
|Collection Tubes with Caps||50 Units|
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