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BugBuster Protein Extraction Reagents
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BugBuster® Protein Extraction Reagent is formulated to gently disrupt the cell wall of E. coli to 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 caused by the release of chromosomal DNA. The clarified extract is ready to use and fully compatible with 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 affinity resin, excess BugBuster Reagent is easily removed by washing the column with the appropriate buffer. BugBuster Reagent 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, BugBuster Reagents and BugBuster plus Benzonase® Nuclease.
The standard BugBuster reagent is supplied as a Tris-buffered “1X”ready-to-use liquid that is stable at room temperature. The 500-ml size is also available with 10 KU Benzonase Nuclease (provided in a separate vial) for the preparation of low viscosity extracts by removal of nucleic acids from protein preparations. BugBuster and Benzonase
are compatible with common protease inhibitors.
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