NOVAGEN BugBaster Bacterial Protein Extraction

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BugBuster Protein Extraction Reagent is formulated for the gentle disruption of the cell wall ofE. coli, resulting in the liberation of soluble protein. It provides a simple, rapid, low costalternative to mechanical methods such as French Press or sonication for releasing expressedtarget proteins in preparation for functional studies or purification. The proprietary formulationutilizes a mixture of non-ionic detergents that is capable of cell wall perforation withoutdenaturing soluble protein.BugBuster (primary amine-free) is intended for applications that are sensitive to the presence ofprimary amines, such as protein immobilization and crosslinking. BugBuster HT is a ready-to-use,premixed formulation of BugBuster and Benzonase® Nuclease ideally suited to high-throughputapplications. BugBuster is also available as a 10X stock that can be diluted in the buffer system ofyour choice, subject to certain considerations (see below for details).Benzonase is a genetically engineered endonuclease from Serratia marcescens (1, 2). Thispromiscuous endonuclease attacks and degrades all forms of DNA and RNA (single stranded,double stranded, linear and circular) and is effective over a wide range of operating conditions(3). The enzyme completely digests nucleic acids to 5′-monophosphate terminatedoligonucleotides 2–5 bases in length (2, 4). Benzonase is ideal for a wide variety of applicationswhere complete digestion of nucleic acids is desirable.BugBuster Protein Extraction Reagent plus Benzonase Nuclease is an efficient combination forgently releasing target proteins and markedly reducing extract viscosity prior to downstreamprocessing. Cells are harvested by centrifugation as usual, followed by suspension in BugBusterreagent at room temperature and Benzonase treatment. During a brief incubation, solubleproteins are released and nucleic acids digested. Insoluble proteins and cell debris are easilyremoved by centrifugation. The resulting low viscosity clarified extract contains soluble proteinsready for additional purification or analysis. The soluble fraction is directly compatible with mostof the chromatography resins offered by Novagen. The insoluble fraction can be furtherprocessed to yield purified inclusion bodies

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