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Fig. 5. Detection of PKc isoform immunoreactivity in oocyte fractions with or without exposure to PMA. (A) Single stage VI oocytes were fractionated into supernatant (Sup) and pellet (Pel) fractions as in Fig. 2 before (-PMA) or after treatment with PMA (1 µM for 30 minutes, +PMA). These fractions were tested for the presence of immunoreactivity for PKC isoforms using isoform-specific antibodies. The approximate mass (in kDa) of each isoform was: delta (75); epsilon (80); eta (76); mu (105); theta (79). Preadsorption of the primary antibody with peptide immunogen abolished these bands. (B) PKC{gamma} immunoreactivity in Xenopus oocytes and the brain and effect of antibody preadsorption. The results show the immunoreactive bands that are detected in supernatant (sup) and pellet (pel) fractions of oocytes probed with PKC{gamma} antibody with or without preadsorption of the antibody with peptide immunogen. Immunoreactivity in a sample of extract (approximately 25 µg protein) from Xenopus brain is shown as a control.