Minute™ Nuclear Envelope Protein Extraction Kit (50 Preps)

Manual & Protocol | Material Safety Data Sheets (MSDS)

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).

Kit includes:

1. Frias A. et al. (2016). A Switch in Akt Isoforms Is Required for Notch-Induced Snail1 Expression and Protection from Cell Death. Mol Cell Biol. 2016 Mar 15; 36(6): 913–922. Prepublished online 2015 Dec 28. Published online 2016 Mar 1. doi: 10.1128/MCB.00842-15.
   
2. Selby, T. L., Biel, N., Varn, M., Patel, S., Patel, A., Hilding, L., ... & Lowrey, A. J. (2019). The Epstein-Barr Virus Oncoprotein, LMP1, Regulates the Function of SENP2, a SUMO-protease. Scientific Reports, 9(1), 9523.
3. Shao, Y., Li, L., Liu, L., Yang, Y., Huang, J., Ji, D., ... & Sun, B. (2022). CD44/ERM/F‐actin complex mediates targeted nuclear degranulation and excessive neutrophil extracellular trap formation during sepsis. Journal of Cellular and Molecular Medicine.

4.  Zervopoulos, S. D., Boukouris, A. E., Saleme, B., Haromy, A., Tejay, S., Sutendra, G., & Michelakis, E. D. (2022). MFN2-driven mitochondria-to-nucleus tethering allows a non-canonical nuclear entry pathway of the mitochondrial pyruvate dehydrogenase complex. Molecular Cell, 82(5), 1066-1077.

5. Kovacs, M. T., Vallette, M., Wiertsema, P., Dingli, F., Loew, D., de Freitas Nader, G. P., ... & Ceccaldi, R. (2023). DNA damage induces nuclear envelope rupture through ATR-mediated phosphorylation of lamin A/C. Molecular Cell.
6. Zhou, X. Y., Lin, B., Chen, W., Cao, R. Q., Guo, Y., Said, A., ... & Zhu, Y. M. (2023). The brain protection of MLKL inhibitor necrosulfonamide against focal ischemia/reperfusion injury associating with blocking the nucleus and nuclear envelope translocation of MLKL and RIP3K. Frontiers in Pharmacology, 14.
7. Taga, K., & Takeuchi, H. (2024). Novel role of host protein SLC25A42 in the HIV‐1 reactivation of latent HIV‐1 provirus. Microbiology and Immunology.