Radiolabeling Cell Tissue Cultures

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.

Was this article helpful?

0 0

Post a comment