The MicroRotofor cell uses liquid-phase IEF to fractionate small volumes (2.5 ml) of proteins in free solution by their pI. The MicroRotofor cell can be used to reduce the complexity of samples prior to any protein analysis technique (including mass spectrometry and 2-D electrophoresis). In 2-D electrophoresis applications, protein fractionation by pI improves downstream sample loading and separation on narrow- and micro-range IPG strips. Whether used alone or as part of a broader separation or purification scheme, the MicroRotofor cell offers rapid, simple, and effective fractionation and enrichment of proteins from complex protein mixtures.
The MicroRotofor cell is easy to set up and offers:
- Unique liquid-phase fractionation by pI, eliminating proteins outside a pI range of interest
- Up to tenfold concentrations of proteins of interest
- Customizable pH gradients — wide or narrow linear pH gradients can be generated using Bio-Lyte® ampholytes
- Rapid recovery of proteins focused into 10 liquid fractions
- Integrated temperature regulation to improve reproducibility and to preserve protein structure and function for native separations
- Preassembled disposable focusing chambers
Accessory equipment required to run the MicroRotofor cell includes a 1,000 V power supply such as the PowerPac™ HV power supply (which is capable of running at 1 W constant power) 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
|Rotofor Cell||Mini Rotofor Cell||MicroRotofor Cell|
|Number of fractions||20||20||10|
|Sample volume||35–60 ml||18 ml||2.5 ml|
|Fraction volume||1.75–3 ml||0.7 ml||200–250 µl|
|Sample load||Milligram to gram||Microgram to milligram||Up to to 1 mg|
MicroRotofor cell lysis kits provide convenient cell lysis and protein extraction protocols to generate total protein samples that are ready for fractionation with a MicroRotofor IEF cell or other downstream applications such as SDS-PAGE and 2-D gel electrophoresis. Kits are available for mammalian, bacterial, plant, and yeast cells.
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