Keating:TEV cleavage

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Protocol for TEV cleavage

• written by Nora

cleaves proteins at specific sequence E-X-X-Y-X-Q-G/S between Q and G/S, with best sequence ENLYFQG

buffer

• 50 mM Tris-HCl pH 8.0
• 0.5 mM EDTA
• 1 mM DTT
• can tolerate up to 100 mM NaCl

cleavage conditions

• optimize according to sample, but here are starting conditions that have worked
• dialyze/resuspend/elute protein sample from 1L culture in 10-25ml TEV buffer
• add 0.5-1 mg TEV
• incubate for 1-3 hr at RT or 3 hr – overnight at 4C