Chromatography (Part 2 of 2): GST Tagging

A simple introduction to the idea of protein tagging and purification of a tagged protein on an affinity chromatography column. The example used is GST tagging followed by purification on a glutathione (GSH) column.
This video was made in association with Trinity College Dublin and was funded by the National Forum for the Enhancement of Teaching and Learning in Higher Education.

[@bikinibottom3864]

This helps a lot! Thanks!

[@younes5126]

Does Thrombine have the same effect as the high concentration of GSH to wash the GST bound protein away ?

[@louisemcateerbiology4855]

Not exactly. Thrombin and concentrated GSH would both cause the target protein to elute from the column, yes. However, thrombin is a protease that cuts the GST tag off the tagged protein. So if thrombin is used, the GST tag remains behind, stuck onto the column. If concentrated GSH is used, the GST tag remains attached to the target protein when it elutes.