Illustrate Column Chromatography Fundamentals

Most biological molecules have a net charge within a pH range of 2 to 10. When the pH is altered, the net charge on molecules can change drastically. In this experiment, proteins are absorbed onto a charged solid support ion-exchange column and subsequently separated during elution using a two-step salt gradient. Students learn basic principals of column chromatography and practice techniques of packing a column, eluting and collecting column fractions, and performing analysis of column fractions through spectrophotometry. Includes materials for six setups.