Keating:TEV cleavage: Difference between revisions
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== Protocol for TEV cleavage == | == Protocol for TEV cleavage == | ||
written by | *written by [[user:ziz|Nora]] | ||
cleaves proteins at specific sequence E-X-X-Y-X-Q-G/S between Q and G/S, with best sequence ENLYFQG | cleaves proteins at specific sequence E-X-X-Y-X-Q-G/S between Q and G/S, with best sequence ENLYFQG | ||
Latest revision as of 12:29, 27 December 2005
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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
