TEV protease cleave protocol

Imidazole doesn't affect TEV efficiency (PDO lab notebook, top of p.81)

Want my protein at 1mg/ml or less, so dilute.

TEV protease wants, ideally, 50mM Tris pH 8.0, 1.5mM EDTA, 3mM DTT. Apparently it's 50% active in 500mM NaCl, too.

Then add 1mg TEV per 10mg my protein (original protocol suggested 1:30 ratio but I had MUCH better results with 1:10 ... I think it's an optimization matter dependent on the batch of TEV you're using-morgan)

Incubate overnight at room temperature (other protocols suggest 3hr at 37 degress, overnight at 4degrees... see which works best for you...). For AF2081, TEV works markedly better at room temperature (Morgan lab notebook #4, p. 70).

Then run straight on column (as yet to be determined whether or not the EDTA is a problem here...)