Immunoblots are developed using Western Lightning enhanced chemilumi-nescence (ECL) reagents (PerkinElmer Life Sciences, Boston, MA). The blot is usually probed by PKA-RI MAb first; this MAb detects both RIa and RIß subunits. To detect the presence of RIIa and RIIß, the blot is first stripped with 20 mM glycine (pH 2.0) solution at 50°C in a microhybridization oven for 30 min, followed by three washings with TBS-T. The remainder of the procedure is as described in Subheading 3.3. A representative blot is shown in Fig. 4, demonstrating the presence of RIa- and RIß-subunit proteins in fraction 1 and RIIa- and RIIß-subunit proteins in fraction 2.
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