Description The nuclear envelope is a very complex membrane-protein system that is notoriously difficult to isolate and purify due to its connection to nuclear and cytoplasmic components. Traditional method of nuclear envelope isolation and purification requires a large amount of starting material and a lengthy and tedious procedure. Invent Biotechnologies MinuteTM 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 ultra centrifugation. Due to the use of protein extraction filter cartridges, the nuclear envelope protein isolation is simple, easy and user friendly. The nuclear envelope proteins are significantly enriched in the final prep. Unlike traditional method that requires large amount 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 min with a final yield of 10-50 µg protein/sample.
Applications The kit is designed to rapidly isolate native nuclear envelope proteins from cultured cells or cells isolated from tissues for applications such as SDS-PAGE, immunoblottings, ELISA, IP, protein translocation analysis, enzyme activity assays and other applications. This kit provides the most rapid method currently available for enrichment of native nuclear envelope proteins.
Buffer Formulations: Proprietary
Kit components (50 preps) 1. 25 ml buffer A 2. 15 ml buffer B 3. 50 protein extraction filter cartridges 4. 50 collection tubes with cap
Storage: Store Buffer A and Buffer B at -20oC upon arrival.
Additional Materials Required 1 X PBS Vortexer Table-Top Microcentrifuge

Important Information: 1. Read the entire procedures carefully. Thaw buffer A and buffer B completely, invert the bottles a few times and place them on ice. Chill protein extraction filter cartridge with collection tube on ice prior to use. 2. All centrifugation steps should be performed at 4oC in a cold room or in a refrigerated mirocentrifuge. 3. To study protein phosphorylation, phosphatase inhibitors (such as PhosStop from Roche) should be added to buffer A prior to use. The use of protease inhibitor cocktails is optional. 4. It is recommended to use BCA Protein Assay Kit for determination of protein concentration (Pierce, Cat #:23227)