The Rotofor and Mini Rotofor cells use liquid-phase IEF to separate proteins into 20 fractions in free solution. As an initial purification step, these cells concentrate proteins and eliminate those outside of a pI range of interest. As a final purification step, they eliminate protein contaminants that may be difficult to remove by other means.
Because it can process up to gram quantities of protein and effectively concentrates proteins into discrete zones based on pI, the Rotofor is a proven and effective method for the enrichment of low-abundance proteins. In a single run, the Rotofor cell concentrates samples up to 20-fold; selected fractions may also be pooled and refractionated on either the Mini Rotofor cell or the MicroRotofor™ cell to yield even greater purification. In 2-D electrophoresis applications, protein fractionation by pI improves downstream sample loading and separation on narrow- and micro-range IPG strips by eliminating proteins outside the pI region of interest.
Accessory equipment required to run the Rotofor and Mini Rotofor cells includes a 3,000 V power supply such as the PowerPac™ HV power supply, a recirculating water chiller, and house vacuum. Bio-Lyte® ampholytes are required to form the internal pH gradient.
Different configurations of the Rotofor system are available to accommodate different sample loads. The Rotofor, Mini Rotofor, and MicroRotofor cells all share the following features:
- Interchangeable cylindrical focusing chambers, which enable refractionation of selected fractions for even greater purification
- Maintenance of biological activity
- Customizable pH gradients — wide or narrow linear pH gradients can be generated using Bio-Lyte ampholytes
- Focusing and enrichment of a protein of interest into 1 or 2 fractions in a single step
||Mini Rotofor Cell
|Number of fractions
||Milligram to gram
||Microgram to milligram
||Up to to 1 mg