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Purification of GST-tagged fusion proteins Glutathione affinity purification, like many other types of affinity purifications, involves three key steps: 1) binding the target protein to the beads, 2) washing away contaminating proteins, and 3) eluting your protein of interest (Figure 3)

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Reduced glutathione with sulfurs lone pair electrons displayed by dots

So, for example, 8M urea or 6M guanidine HCl would likely denature GST, preventing the GST-glutathione interaction that serves as the foundation for this purification