Optimization of a Reverse Phase High Performance Liquid Chromatography Pre-fractionation Method Followed By Two Dimensional Electrophoresis

Prefractionation of comples protein mixtures is an efficient method in orther to increase the separation power of two - dimensional electrophoresis (2DE). Reverse Phase High Performance Liquid Chromatography (RP- HPLC) has been successfully utilized for prefractionation followed by 2DE. Here, optimization of a RP-HPLC prefractionation method of BHK cells lysate followed by 2DE is reported. A step gradient elution of cell lysate was improved to yield properly resolved fractions for subsequent analysis by 2DE.