Using Ion Exchange Chromatography to Separate Proteins (Lab 9C)

This kit is designed to teach 8 groups of four students the principles of Ion Exchange Chromatography and protein purification.  The kit contains the reagents to allow students to:

• Equilibrate a DEAE agarose column
• Load a protein sample on the column (amylase, lysozyme or a mixture of both) 
• Wash the unbound protein sample from the column
• Bind and then elute the bound protein sample from the column with a salt buffer
• Measure the amount of eluted protein with a Bradford protein assay
• Generate an elution profile determining which protein blinded to the column and which flowed through
• Give evidence that the two proteins were separated by the chromatography technology

Features

• For 8 groups of 4 students 
• Requires micropipettes and tips
• Optional requirement for a spectrophotometer

Applications

• HS-LS1: From Molecules to Organisms: Structures and Processes
• LS1.A: Structure and Function