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. Author manuscript; available in PMC: 2010 Oct 13.
Published in final edited form as: Biochem J. 2009 Dec 10;424(3):399–410. doi: 10.1042/BJ20090842

Figure 4. Insulin- or CL-induced assembly of macromolecular complex(es) containing PDE3B and signalling proteins: co-immunoprecipitation with PDE3B.

Figure 4

Superose 6 fractions, as in Figure 3, were divided into three groups (A, B and C), and pooled for co-immunoprecipitation (Group A, fractions 17–21; Group B, fractions 22, 24, 26, 28 and 30; Group C, fractions 32, 34, 36, 38 and 40). Pooled fractions were cleared, and complexes containing PDE3B were immunoprecipitated with 20 μl of anti-PDE3B-CT antibodies. Group A fractions were also immunoprecipitated with control rabbit IgG. Protein G–Sepharose-bound proteins were eluted in 100 μl of Laemmli’s sample buffer. Samples (15 μl) were subjected to SDS/PAGE and immunoblotting with specific antibodies as indicated. One representative experiment is shown (n = 3), with the exception that immunoprecipitation/Western blot analysis for β3-AR were conducted in two experiments. IP, immunoprecipitation.