EMD Millipore Develops Simplified Protocol for Rapid Purification of Recombinant Proteins

Benzonase Nuclease - Effective viscosity reduction and removal of nucleic acids from protein solutions Benzonase Nuclease is a genetically engineered endonuclease from Serratia marcescens. It degrades all forms of DNA and RNA (single stranded, double stranded, linear and circular) while having no proteolytic activity. Benzonase Nuclease is effective over a wide range of conditions and possesses an exceptionally high specific activity. The Benzonase Nuclease enzyme completely digests nucleic acids to 5′-monophosphate terminated oligonucleotides 2 to 5 bases in length (below the hybridization limit)Merck Benzonase Nuclease is ideal for removal of nucleic acids from recombinant proteins, enabling compliance with FDA guidelines for nucleic acid contamination. The ability of Benzonase to rapidly hydrolyze nucleic acids makes the enzyme an excellent choice for viscosity reduction to reduce processing time and increase yields of protein. For example, the enzyme is compatible with BugBuster® and PopCulture® Protein Extraction Reagents and can therefore be added along with these reagents to eliminate viscosity and remove nucleic acids from E. coli extracts.The Benzonase Nuclease  enzyme consists of two subunits of 30 kDa each. It is functional between pH 6 and 10 and from 0°C to 42°C and requires 1–2 mM Mg2+ for activation. The Merck Benzonase Nuclease enzyme is also active in the presence of ionic and non-ionic detergents, reducing agents, PMSF (1 mM), EDTA (1 mM) and urea (relative activity depends on specific conditions). Activity of Benzonase Nuclease is inhibited by > 150 mM monovalent cations, > 100 mM phosphate, > 100 mM ammonium sulfate, or > 100 mM guanidine HCl. Benzonase Nuclease is available in ultrapure (> 99% by SDS-PAGE) and pure (> 90%) grades. Both preparations are free of detectable protease and have specific activity > 1 × 106 units/mg protein. The > 99% purity grade of Merck Benzonase Nuclease is tested for endotoxins and contains < 0.25 EU/1,000 units. Supplied as a 0.2 µ filtered solution in 50% glycerol. Store at -20°C.

BugBuster® Master Mix combines BugBuster Protein Extraction Reagent with Benzonase® Nuclease and rLysozyme™ Solution in one convenient reagent. BugBuster Master Mix allows for maximum recovery of active soluble protein from both Gram-negative and Gram-positive bacteria. With the Master Mix, there is no need for dilution or separate addition steps. The two available package sizes provide sufficient reagents for protein extraction from 20 g and 100 g cell paste.

Magnetic Bead-Based Isolation of Proteins Millipore’s new PureProteome magnetic beads ensure the rapid and reproducible isolation of proteins. Unlike conventional methods that require centrifugation to pellet the beads then careful aspiration to avoid sample loss, PureProteome magnetic beads are isolated using a magnetic rack. This allows for the total removal of buffers for complete recovery of beads no sample dilution. You eliminate variability while maximizing recovery.Customize your own bead with PureProteome's NHS FlexiBind magnetic bead. NHS FlexiBind gives you the flexibility to bind the ligand of your choice! Add your own ligand in less than 60 minutes, the only requirement is that your ligand must contain a free primary Amine group.PureProteome™ Magnetic Beads Advantages: High capacity - Advanced chemistry combined with high surface area provides more binding sites for proteins. Consistent results with no sample loss - Particles visible as they adhere to side of tube for quick and easy aspiration and complete buffer removal Fast processing time - Beads are immobilized in seconds. Increased kinetics of bead-protein binding enables shorter incubations Economical - Significantly more affordable then competitive magnetic beads