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BugBuster® Protein Extraction Reagents
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Simple extraction of active protein from E. coli without sonication
BugBuster® Protein Extraction Reagent is formulated to gently disrupt the cell wall of E. coli and liberate soluble proteins. It provides a simple, rapid, low-cost alternative to mechanical methods such as French Press or sonication for releasing expressed target protein in preparation for purification or other applications. The proprietary formulation utilizes a detergent mix that is capable of cell wall perforation without denaturing soluble protein.
In practice, cells are harvested by centrifugation and suspended in BugBuster. The addition of rLysozyme™ Solution, which hydrolyzes N-acetylmuramide linkages in the cell wall, enhances the extraction efficiency, especially for larger proteins. Following a brief incubation, insoluble cell debris is removed by centrifugation. At this point, Benzonase® Nuclease can be added to reduce the viscosity of the extract due to liberation of chromosomal DNA. The clarified extract is ready to use and fully compatible with the affinity supports offered by Novagen, including GST•Bind™, GST•Mag™, His•Bind®, His•Mag™, S•Tag™, Strep•Tactin®, and T7•Tag® Resins, or several other chromatography matrices. Following binding to the affinity resin, excess BugBuster is easily removed by washing the column with the appropriate buffer. BugBuster is also useful for the preparation of high-purity inclusion bodies in instances where expressed proteins are insoluble. The reagent is available in a variety of configurations. |
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