Liquid-phase isoelectric focusing on small volumes

Rotofor including micro and mini systems allow the pre-fractionation of protein mixtures for sub-proteome analysis. The device separates complex protein samples in free solution by preparative isoelectric focusing (IEF).

In special experiments protein activity can be retained making Rotofor a preparative tool to purify proteins.

The Rotofor rapidly isolates proteins of interest from the bulk proteins in complex biological samples and provides purified, biologically active proteins for functional and structural studies, antibody production, and sequence analysis.

Ten or less fractions (microRotofor: 200–250 µl/fraction) can be collected from one run, pooled, and also refractionated for further purification. Fractionation by liquid-phase IEF is particularly beneficial for those proteins that are insoluble or otherwise do not separate well in other, gel-based IEF media. Moreover, IEF in solution enables fractionation of proteins in their native state by isoelectric point (pI). Ampholytes are used to generate the pH gradient for separation so that a continuous customizable pH gradient may be formed.

Test with the large Rotofor

Task: Separate colored proteins

phycocyanin, blue, pI 4.5-5hemoglobin, red, pI 7.2-7.6cytochrome c, organge pI 9.5-10
After the run in the Rotofor chambers
After elution into test tubes