The atypical protein kinase C isoform, PKCζ, is critical for mediating mitogenic signal transduction, cell survival, and some of the physiological actions of insulin. PKCζ pseudosubstrate inhibitor is a synthetic peptide that corresponds to a pseudosubstrate domain of this PKC isoform. It selectively, reversibly, and substrate-competitively inhibits PKCζ activity and, thus, is used to delineate the signaling functions of PKCζ.