Using SDS-PAGE to assess the purification of protein

Overview

Teaching: 30 min
Exercises: 180 min

Questions

• How can SDS-PAGE assess the purity of a protein?

Objectives

• Students will understand how acrylamide gels are made and manipulated.

• Students will be able to estimate protein purity using SDS-PAGE.

• Students will characterize the size of a protein using SDS-PAGE.

In this BASIL module, students use Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) as a separation technique to assesses the quality of a protein purification scheme. The approach of this module is to explain the mechanism by which SDS-PAGE is able to separate proteins and demonstrate the importance of this technique by having students run their purification samples on a gel. Students will be required to use SDS-PAGE to determine the purity of their protein samples, evaluate the size of their protein product, and correlate their experimental data with known information regarding the size of their protein.

Module Resources

Download student module here

Key Points

• To carry out this lab students need vertical electrophoresis equipment.

• The students can use pre-purchased gels or pour their own.