ReadyPreps™ Protein Preparation Kit

Note: This product will be discontinued when current inventories are sold out.

Applications

• Rapid screening of large numbers of expression clones for desired protein activity, for testing the effects of various parameters (such as culture conditions and induction protocols) on the yield of active protein.
• Simultaneous isolation of plasmid and expression product from the same preparation.
• Preparative-scale protein purification.

The ReadyPreps™ Protein Preparation Kit is a rapid, gentle, enzymatic method for lysing bacterial cells and reducing viscosity for the extraction of proteins, especially recombinant gene products expressed in E. coli.¹ The kit uses Ready-Lyse™ Lysozyme Solution, a stable nonavian, nonmammalian lysozyme that has a higher specific activity than egg-white lysozyme. The higher specific activity of Ready-Lyse Lysozyme allows the addition of less lysozyme compared to egg-white lysozyme, minimizing the amount of extraneous protein in the preparation. OmniCleave™ Endonuclease reduces the viscosity of cell lysates, facilitating handling and processing. RNase I is also provided for removing cellular RNA when the plasmid and protein are extracted simultaneously. The nucleases are included separately and can be added as needed.

The ReadyPreps Kit can be used to rapidly prepare small samples for in-process monitoring of expression, for screening large numbers of transformants for a desired activity, or for larger scale production of proteins. In addition, it is possible to extract protein and plasmid DNA simultaneously from the same sample, which may be important in mutagenesis and library screening. By assaying the resulting lysates for activity, the ReadyPreps Kit enables a quick determination of the production of functional proteins.

Benefits

• Most preparations can be completed in 20-45 minutes, reducing exposure of proteins to cellular proteases.
• Process large numbers of samples simultaneously.
• No heating or foaming of the lysate that can denature proteins as is possible with some mechanical lysis methods.
• More reproducible than mechanical lysis methods.
• Use with single colonies or milligram quantities of cells.
• Can be configured for nucleic acid-free protein preparations or for simultaneous protein and plasmid isolation.
• Compatible with most downstream purification processes including DNA precipitation methods (e.g., PEG or PEI), ammonium sulfate precipitation, anion exchange, affinity (e.g., histidine-tagged, maltose-binding protein) chromatography, and tangential flow filtration.

Reference

1. Schoenfeld, T. (1997) EPICENTRE Forum 4(3), 7.