Sauer:Expression/purification of 35S-Met proteins
- In general, the Sauer lab only radiolabels His6-tagged proteins because they are easy to purify without getting radiation all over the place. If necessary, however, other methods of purification can be employed, but it is recommended that gravity columns be used versus an FPLC-type system in order to reduce the spread of radiation.
- In general, either of these protocols should work for your protein of choice. Major differences include the time of induction with and without radiolabel, and native versus denaturing protein purification by nickel column affinity.