Inclusion body preparation and a 96-well matrix of buffers and additives for convenient optimization of refolding conditions Protein functional and structural studies often require a large amount of pure, correctly folded protein, which is commonly produced in Escherichia coli (E. coli) expression systems. However, production of foreign proteins in E. coli can result in the formation of inclusion bodies (IB) — dense, insoluble, aggregates of misfolded protein. IB also have useful attributes — they are easily purified, resistant to proteolysis, and can be solubilized with chaotropic agents. Defining conditions that promote refolding of a chemically denatured protein into its native conformation is empirical and often time consuming. Simultaneous and systematic evaluation of a large number of refolding conditions increases chances of identifying an optimal refolding condition for a given protein. To take the tedium and guesswork out of finding the optimal refolding condition for your target protein, iFOLD® Protein Refolding Systems provide comprehensive refolding screens in a 96-well plate format. These systems are based on an extensive literature review of successful refolding experiments and information contained in the REFOLD database (http://refold.med.monash.edu.au). The systems differ in the chemistry used to denature the inclusion bodies and in the refolding additives included in the 96-well plate. In addition to a 96-well plate containing 92 (System 1), 94 (System 2) or 91 (System 3) unique buffer and refolding additive combinations, the systems include the reagents needed to purify inclusion bodies and solubilize the component proteins. Significantly, all steps of the refolding screens are equally compatible with manual and high-throughput automated liquid handling. |
 | Features:
- Cover nearly 300 refolding conditions
- Include all reagents for inclusion body purification
- Include pre-dispensed 96-well plate-based refolding buffer matrix
- Test pH range 7.0 – 9.0
- Refolding additives include salts, cyclodextrin, redox agents, chaotropes, glycols, nondetergent sulfobetaines (NDSBs), and unique FoldACE™ reagents
- Refolding conditions based on extensive literature review and REFOLD database
- Compatible with high-throughput automated liquid handling
- Amenable to large-scale protein refolding based on the identified positive refolding buffer conditions
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