1. Grow cells in 6-well plate. If cells are transfected, label cells 24-48 hours after
transfection.
2. Rinse cells twice with methionine and cystine free growth media.
3. (optional) Add 1.5 ml methionine and cystine free growth media supplemented
with 10% dialyzed FCS. Culture 4-16 hours.
4. Add 20-30 ml of Expre35S35S ( 35S-Express )protein labeling mix (NEN).
Label cells at 37 °C for 1-2 hours.
5. Add 1.5 ml normal media with methionine, cystine, and 10% serum. Chase 4 hour to over night at 37 °C.
6. Remove media, and wash twice with PBS, and harvest cells.
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