Campylobacter jejuni cells were grown to mid-log phase in a defined broth, and the samples harvested. In both cases, cells were radiolabelled with 15 mCi/ml [35S]-methionine [ICN] for 30 mins. The first dimension was run using 11 cm isoelectric focussing strips [Pharmacia; linear 3-10] for a total of 24300 Vh, and ~ 1 mg of protein was loaded. The second dimension used 12% resolving gels run at 15 mA per gel overnight.
(a) The above gel was silver stained and dried before putting down to autoradiograph for 30 days.
(b) This gel was electroblotted onto nitrocellulose membranes (100 volts for 1 hour) before drying and prepared using the EA-wax system. The gel was then put down to autoradiograph for 5 days.
Back to Technical Evaluation Page