Many transcription factors are regulated by post-translational phosphorylation mechanisms. This chapter described several approaches that have been utilized to examine the phosphorylation of the c-Jun transcription factor. A combination of the techniques described in this chapter can be used to determine whether a transcription factor is phosphorylated, in vivo, to analyze the sites of phosphorylation in vitro, and to permit the identification of putative protein kinases that may mediate this phosphorylation in vivo. Proteins labeled with 32P by either in vivo or in vitro techniques can be further analyzed by tryptic phosphopeptide mapping or phosphoamino acid analysis. These procedures have been described in detail elsewhere.
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