1. The cultures or tissue explants are rinsed in culture medium missing the appropriate substrate. Thus, the medium must be methionine-free or phosphate-free if these are to be the radiolabeled compounds.
2. The cultures are then incubated in the same medium containing 1 mCi/mL of the radiolabeled compound, at 37°C, in tissue-culture conditions for 2-3 h.
3. The radiolabeled medium is removed, and the embryonic tissue is washed once in "cold" medium before being dissociated in either 2X dissociation buffer or RIPA/NP40 lysis buffer.
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