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IN VIVO SYSTEMS > TISSUE CULTURE > 35S Labeling > 35S Methionine Labeling

Labeling cells with 35S methionine

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You will need

  1. DMEM minus methionine (Gibco-BRL) with glutamine and pen-strep added
  2. dialysed FBS (dialyse against PBS for several hours)
  3. 35S methionine

For one 100mm dish of cells

A. To label overnight:

  1. remove medium, add 5ml DMEM - met, incubate cells for 30min
  2. remove and add 5ml DMEM - met + 10% dialysed FBS + 0.25mCi 35Smet incubate 16hours. Be sure to label dishes with radioactive tape!
  3. remove medium and prepare an NP40 lysate for immunoprecipitation.

B. To pulse label for 2hrs:

  1. remove medium, add 5ml DMEM - met, incubate cells for 30min
  2. remove and add 5ml DMEM - met + 0.25 to 1mCi 35Smet, incubate for 2hours
  3. remove medium and prepare an NP40 lysate for immunoprecipitation.

Dispose of all radioactive waste appropriately!