Keating:TEV cleavage

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Protocol for TEV cleavage

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
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