Size Exclusion Chromatography Lab Report

1. Background
MCB 253
Evan London
2/5/16
A Comparison of
Size Exclusion and Affinity Chromatography In molecular cell biology experiments, accuracy is one of the most important factors in obtaining reproducible results. A luxury not afforded to molecular cell biologists is the ability to easily manipulate samples since the material they deal with is at a microscopic level. Proteins represent a unique opportunity for study because of their intrinsic relation to the central dogma of biology. However proteins are just one subset of macromolecules found in organisms and each protein has its own unique structure that determines its function. When a sample is collected for use in an experiment it is therefore necessary to separate the protein of interest from other macromolecules along with undesired proteins using a series of mechanical and chemical processes. These process of protein purification takes many forms and most rely on the principles of chromatography.

2. Technique Descriptions

2.1 Size Exclusion Chromatography (SEC) As the name
In an affinity chromatography experiment the separation of the sample is based on the binding of the solute to binding sites present within the matrix of the stationary phase. Proteins especially lend themselves to being purified through this technique because of the information we know about amino-acid sequences and the various motifs that can be found within proteins. An application of affinity chromatography for protein purification which is very effective is immunoprecipitation. In this technique specific antibodies are attached to the matrix and are used to capture protein targets. Immunoprecipitation has applications for identifying protein modifications and even pathway mapping (Uhlén, 2008). The flexibility of this technique definitely is one of its major