Size Exclusion Chromatography Lab Report

The purpose of this experiment is to analyze and identify specific proteins from a mixture of proteins using the purify technique of size exclusion chromatography. The use of size exclusion chromatography also called Gel filtration allows to separate proteins according to size (molecular weight) and shape. The idea of separating molecules base of the physical sizes depends on the stationary phase and the mobile phase. Selecting the appropriate stationary phase and mobile phase is vital to obtain successful separations. Stationary phase is made of microscopic beads which consist of a cross-linked polysaccharide which forms porous beads or bead-like matrix that is packed into a column. Mobile phase is a solvent, normally consists of a buffer,

Smaller molecules can diffuse into the beads and move more slowly down the column. The process of extracting one material from another by washing with a solvent is called elution; they are three ways isocratic elution, gradient elution, and stepwise elution. In this experiment the process of isocratic elution will be uses, where the same mobile phase is used throughout the elution and allows separation of molecules that differ significantly in size. By varying the degree of cross-linking the gels are optimized for different molecular weight ranges.The purpose of this experiment is to analyze and identify specific proteins from a mixture of proteins using the purify technique of size exclusion chromatography. The use of size exclusion chromatography also called Gel filtration allows to separate proteins according to size (molecular weight) and shape. The idea of separating molecules base of the physical sizes depends on the stationary phase and the mobile phase. Selecting the appropriate stationary phase and mobile phase is vital to obtain successful