Macromolecules Lab Report

The human body produces various essential proteins that provide functionality in the biochemistry of our system. How individual molecules are separated or a mixture of molecules are separated can be dependant of their size. Size exclusion chromatography (SEC), is the controlled separation technique that is based on the size of the particles in relation to the pore size (Barth and Saunders 2012). The separation process veers to be quick for large molecules and slower for smaller molecules. The process would involve the large macromolecules to permit through the column bed, as the smaller molecules infiltrated the pores. Molecules passing through the column bed are divided between the stationary and mobile phases, and these molecules would be …show more content…
The remaining 10 tubes were number from 1- 10 and all collection tubes were placed in a test tube rack. 4 ml of column buffer was added to the test tube marked ‘column buffer’ using a pipette. A sizing chromatography column was obtained with the lid and the end snapped off and placed in the ‘waste’ test tube. A column cap was placed at the bottom of the sizing column, once the buffer had drained completely within the ‘waste’ tube. The sizing chromatography column was then placed into column 1 and was loaded with the protein sample. The protein mix was added just above the column bed for minimal disturbance. 250 µl were added to the column buffer placed within test tube 1. Column buffer proceeded to run through the column bed and drops were collected within test tube 1. Another 250 µl of column buffer were added to the column bed where more drops were collected in tube 1. 3 ml of column buffer were added to the column matrix using a pipette and the sizing column was transferred to test tube 2. 5 drops of buffer were counted and collected in each test tube from test tube 2 to 9, until test tube 10, where 10 drops of buffer were collected and the sizing column was